pCT74-HygR-RFP

LOCUS       pCT74-HygR-RFP                5056 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 5056)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2024 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..35
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             434..1459
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(1500..2195)
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     promoter        complement(2213..2231)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2252..2268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2276..2292
                     /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(2300..2330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2345..2366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2654..3242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3413..4273)
                     /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(4274..4378)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4404..4859)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5001..5017
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5024..5042
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga attcatcgat
       61 accgtcgacg ttaactgata ttgaaggagc attttttggg cttggctgga gctagtggag
      121 gtcaacaatg aatgcctatt ttggtttagt cgtccaggcg gtgagcacaa aatttgtgtc
      181 gtttgacaag atggttcatt taggcaactg gtcagatcag ccccacttgt agcagtagcg
      241 gcggcgctcg aagtgtgact cttattagca gacaggaacg aggacattat tatcatctgc
      301 tgcttggtgc acgataactt ggtgcgtttg tcaagcaagg taagtggacg acccggtcat
      361 accttcttaa gttcgccctt cctcccttta tttcagattc aatctgactt acctattcta
      421 cccaagcatc caaatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
      481 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
      541 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
      601 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
      661 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
      721 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctcc agccggtcgc
      781 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
      841 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
      901 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
      961 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     1021 tgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     1081 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatcc tcttctggag
     1141 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     1201 tgcaggatcg ccgcgcctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     1261 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     1321 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     1381 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     1441 tccgagggca aaggaataga gtagatgccg accgggaacc agttaacgtc gacctcgagt
     1501 catctgtgcc ccagtttgct agggaggtcg cagtacttgg ccacagccat ctcgtgctgc
     1561 tcgacgtagg tctctttgtc ggcctccttg attctttcca gtctgtggtc cacgaagtgg
     1621 aagccgggca tcttgaggtt cttagcgggt ttcttggatc tgtatgtggt cttgaaggag
     1681 cagtgcaggt agcccccgcc cacgagcttc agggccatct ggctgtggcc tctcaggccg
     1741 ccgtcagcgg ggtacagcat ctcggtgttg gcctcccagc cgcgtgtttt cttctgcatc
     1801 acagggccgt tggatgggaa gttcaccccg ttgatcttga cgttgtagat gatgcagccg
     1861 ttctggaagc tggtgtcctg ggtagcggtc agcacgcccc cgtcttcgta tgtggtgatt
     1921 ctctcccatg tgaagccctc agggaaggac tgcttaaaga agtcggggat gccctgggtg
     1981 tggttgatga aggctttgct gccgtacatg aagctggtag ccaggatgtc gaaggcgaag
     2041 gggagagggc cgccctcgac caccttgatc ttcatggtct gggtgccctc gtagggcttg
     2101 ccttcgccct cggatgtgca cttgaagtgg tggttgttca cggtgccctc catgtacagc
     2161 ttcatgtgca tgttctcctt gatcagctcg ctcatggtac cagcttttgt tccctttagt
     2221 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2401 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2461 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2521 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3061 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3361 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4141 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4201 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4261 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4381 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     4441 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4501 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4561 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4621 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4681 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4741 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4801 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4861 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     4921 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     4981 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
     5041 gggcgaattg gagctc
//