pCT74

LOCUS       pCT74                5790 bp DNA     circular SYN 17-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5790)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2024 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..35
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(78..330)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(349..1068)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             1855..2880
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(2947..2965)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2986..3002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3010..3026
                     /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(3034..3064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3079..3100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3388..3976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4147..5007)
                     /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"
     promoter        complement(5008..5112)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5138..5593)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5735..5751
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5758..5776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga attctcatgt
       61 ttgacagctt atcatcggat ctagtaacat agatgacacc gcgcgcgata atttatccta
      121 gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat
      181 cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac
      241 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa
      301 gaaactttat tgccaaatgt ttgaacgatc tgcagccggg cggccgcttt acttgtacag
      361 ctcgtccatg ccgtgagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
      421 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
      481 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
      541 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
      601 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
      661 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
      721 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
      781 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
      841 ggggtagcgg ctgaagcact gcacgccgta ggtgaaggtg gtcacgaggg tgggccaggg
      901 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
      961 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     1021 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg cctatattca
     1081 ttcattgtca gctatcgctg gcaacaagat ataagcgtgt acttacgatt ctaggaccct
     1141 gcgaatagag ttggagttgg ataccgatgg acgattgtta gataggacgg gatgctgata
     1201 gtcgaggtaa gttgttgagg gtggtggttt tatcaaggac ggacacctta gccctttata
     1261 tacacagaat cttattgcta ggtagccgga cggtgcggat tccaggtatc agtaggccct
     1321 ctgaatgtcc atgcaatgat gcccaacaat cgagacgatc ctacttcgga accgactcga
     1381 tcctcgtcgg gcctacatgt atgagaccat gtgcacctcc gaagcatcct tacggagagc
     1441 ggaattgcgc gtacagaact cctccatgca ttccaatcga taccgtcgac gttaactgat
     1501 attgaaggag cattttttgg gcttggctgg agctagtgga ggtcaacaat gaatgcctat
     1561 tttggtttag tcgtccaggc ggtgagcaca aaatttgtgt cgtttgacaa gatggttcat
     1621 ttaggcaact ggtcagatca gccccacttg tagcagtagc ggcggcgctc gaagtgtgac
     1681 tcttattagc agacaggaac gaggacatta ttatcatctg ctgcttggtg cacgataact
     1741 tggtgcgttt gtcaagcaag gtaagtggac gacccggtca taccttctta agttcgccct
     1801 tcctcccttt atttcagatt caatctgact tacctattct acccaagcat ccaaatgaaa
     1861 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     1921 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     1981 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     2041 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag
     2101 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     2161 ctgcctgaaa ccgaactgcc cgctgttctc cagccggtcg cggaggccat ggatgcgatc
     2221 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     2281 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     2341 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     2401 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc atgcggattt cggctccaac
     2461 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     2521 ggggattccc aatacgaggt cgccaacatc ctcttctgga ggccgtggtt ggcttgtatg
     2581 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcgcctc
     2641 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     2701 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     2761 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     2821 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     2881 agtagatgcc gaccgggaac cagttaacgt cgacctcgag ggggggcccg gtaccagctt
     2941 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     3001 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3061 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3121 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3181 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3241 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3301 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3361 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3421 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3481 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3541 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3601 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3661 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3721 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3781 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3841 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3901 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3961 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4021 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4201 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4261 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4321 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4381 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4441 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4501 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4561 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4621 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4681 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4741 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4801 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4861 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4921 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4981 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5041 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5101 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     5161 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5221 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5281 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5341 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5401 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5461 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5521 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5581 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5641 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     5701 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
     5761 tacgactcac tatagggcga attggagctc
//