Antares-SEPLuc

LOCUS       Antares-SEPLuc                9092 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  precursor of the bioluminescent reporter pHluc, for detecting 
            extracellular pH changes in tumor acidosis.
ACCESSION   .
VERSION     .
KEYWORDS    Antares-SEPLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9092)
  AUTHORS   Ong TT, Ang Z, Verma R, Koean R, Tam JKC, Ding JL
  TITLE     pHLuc, a Ratiometric Luminescent Reporter for in vivo Monitoring of 
            Tumor Acidosis.
  JOURNAL   Front Bioeng Biotechnol. 2020 May 8;8:412. doi: 
            10.3389/fbioe.2020.00412. eCollection 2020.
  PUBMED    32457886
REFERENCE   2  (bases 1 to 9092)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9092
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(23..40)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    29..253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      299..727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(386..405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     596..617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(736..756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        741..1070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      921..1056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     983..1002
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1310..1431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1347..1366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1401..1420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1480..1496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1480..1496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1493..1515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1504..1520
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1528..1558)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1573..1594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1711..1728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1882..2470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1962..1981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2641..3501)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3264..3283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3502..3606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3681..3700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3872..4251
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4252..4455
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4406..4426
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4500..4519
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4500..4518
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6411..6464
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6471..7070
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6471..6490)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6967..6987
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7072..7658
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7239..7256)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7466..7485
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
       61 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
      121 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
      181 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
      241 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
      301 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      361 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      421 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      481 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      541 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      601 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      661 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      721 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
      781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
      841 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      901 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      961 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1021 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1081 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcgcgg
     1141 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     1201 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     1261 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     1321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     1441 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3601 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     3661 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     3721 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     3781 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     3841 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     3901 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3961 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     4021 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     4081 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     4141 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     4201 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     4261 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     4321 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     4381 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     4441 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     4501 aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg
     4561 agctcggatc cactagtcca gtgtggtgga attcatggtg agcaagggcg aggagctgat
     4621 caaggagaac atgagaagca agctgtacct ggaaggcagc gtgaacggcc accagttcaa
     4681 gtgcacccac gaaggggagg gcaagcccta cgagggcaag cagaccaaca ggatcaaggt
     4741 ggtggaggga ggccccctgc cgttcgcatt cgacatcctg gccacccact ttatgtacgg
     4801 gagcaaggtg ttcatcaagt accccgccga cctccccgat tattttaagc agtccttccc
     4861 tgagggcttc acatgggaga gagtcatggt gttcgaagac gggggcgtgc tgaccgccac
     4921 ccaggacacc agcctccagg acggcgagct catctacaac gtcaaggtca gaggggtgaa
     4981 cttcccagcc aacggccccg tgatgcagaa gaaaacactg ggctgggagc ccagcaccga
     5041 gaccatgtac cccgctgacg gcggcctgga aggcagatgc gacaaggccc tgaagctcgt
     5101 gggcgggggc cacctgcacg tcaacttcaa gaccacatac aagtccaaga aacccgtgaa
     5161 gatgcccggc gtccactacg tggaccgcag actggaaaga atcaaggagg ccgacaacga
     5221 gacctacgtc gagcagtacg agcacgctgt ggccagatac tccaacctgg gcggaggctt
     5281 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     5341 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     5401 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     5461 gtatgaagga ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     5521 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     5581 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     5641 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     5701 gcattctggc gaggcacgag ctgatcaagg agaacatgag aagcaagctg tacctggaag
     5761 gcagcgtgaa cggccaccag ttcaagtgca cccacgaagg ggagggcaag ccctacgagg
     5821 gcaagcagac caacaggatc aaggtggtgg agggaggccc cctgccgttc gcattcgaca
     5881 tcctggccac ccactttatg tacgggagca aggtgttcat caagtacccc gccgacctcc
     5941 ccgattattt taagcagtcc ttccctgagg gcttcacatg ggagagagtc atggtgttcg
     6001 aagacggggg cgtgctgacc gccacccagg acaccagcct ccaggacggc gagctcatct
     6061 acaacgtcaa ggtcagaggg gtgaacttcc cagccaacgg ccccgtgatg cagaagaaaa
     6121 cactgggctg ggagcccagc accgagacca tgtaccccgc tgacggcggc ctggaaggca
     6181 gatgcgacaa ggccctgaag ctcgtgggcg ggggccacct gcacgtcaac ttcaagacca
     6241 catacaagtc caagaaaccc gtgaagatgc ccggcgtcca ctacgtggac cgcagactgg
     6301 aaagaatcaa ggaggccgac aacgagacct acgtcgagca gtacgagcac gctgtggcca
     6361 gatactccaa cctgggcgga ggcatggacg agctgtacaa ggcggccgcc gagggcagag
     6421 gaagtcttct aacatgcggt gacgtggagg agaatcccgg cccttccggg atgaccgagt
     6481 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     6541 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     6601 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     6661 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     6721 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     6781 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     6841 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     6901 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     6961 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     7021 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acccctctcc
     7081 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     7141 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     7201 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     7261 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     7321 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     7381 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     7441 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     7501 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     7561 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     7621 tttcctttga aaaacacgat gataatatgg ccacaaccat ggttgagatg ttgccaaccg
     7681 tagctgtgct cgttttggct gtcagcgtag tcgctaaaga taatacaaca cttcaggagt
     7741 tcgcaacatc taagggagaa gagttgttta ctggcgtagt acccattctt gtggaacttg
     7801 acggcgatgt taatggtcat aagttctctg tgtctggtga gggcgagggg gatgctacat
     7861 atggtaaatt gaccctcaaa tttatttgta caaccgggaa gctccctgta ccctggccta
     7921 ctctcgtgac caccctcacc tacggggtac aatgtttttc cagatacccc gatcacatga
     7981 agcgccacga tttcttcaaa agtgccatgc ccgagggtta cgtccaggaa cgcacaatat
     8041 ttttcaagga tgatggaaat tacaaaaccc gcgccgaggt taagttcgag ggggataccc
     8101 tcgttaaccg gattgaactg aagggtatcg actttaagga ggacggtaat atattggggc
     8161 ataaattgga atacaactat aacgaccacc aggtgtatat catggccgat aaacagaaga
     8221 acggcatcaa agcaaacttt aaaattaggc ataacataga ggatggtggc gttcagcttg
     8281 ctgatcacta tcaacagaat acccctatag gtgatggacc cgtgctcctc ccagataacc
     8341 attacttgtt tacaacttct acactctcta aggaccccaa cgaaaagaga gaccacatgg
     8401 tgctcttgga gttcgtaact gccgccggaa ttacacatgg tatggacgag ctctacaagg
     8461 atatttccgg gggggtcttt acacttgaag atttcgtcgg ggattggaga caaactgccg
     8521 gttataatct tgatcaggtg cttgagcaag ggggggtgag ctcccttttt caaaacctgg
     8581 gtgtatctgt cacccctatc caaaggattg tgcttagcgg tgaaaatggt ctcaaaatcg
     8641 acatccacgt gatcatccca tacgagggtc ttagtggaga tcagatgggg cagatagaga
     8701 agattttcaa agttgtttac cctgtcgacg accaccattt caaagtaatt ctgcactacg
     8761 gaaccctcgt catagacggg gtcacaccca acatgataga ttactttggg cgaccctatg
     8821 aagggatagc agtattcgac ggcaaaaaga taaccgtgac aggcaccctg tggaatggaa
     8881 acaaaattat agatgagcgc ctgatcaacc ctgacggttc tttgttgttc agggttacaa
     8941 tcaatggtgt cacaggctgg aggctctgtg agcgcatcct ggctcagttg gagaatgggg
     9001 ggatctccct cttggtgcaa aatacatcat ggatgttgct tcttcttctt agcttgagcc
     9061 tcttgcaggc attggatttc atcagtctct aa
//