LOCUS Exported 8568 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCA-mTmG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8568) TITLE pCA-mTmG REFERENCE 2 (bases 1 to 8568) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8568 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(6..461) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(143..164) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 355..374 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 588..610 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 602..619 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 603..619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 626..645 /label=T7 /note="T7 promoter, forward primer" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 669..685 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 670..686 /label=KS primer /note="common sequencing primer, one of multiple similar variants" enhancer 791..1094 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1096..1373 /label=chicken beta-actin promoter intron 1375..2392 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 2316..2338 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2400..2419 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 2494..2527 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 2570..4000 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" intron 4084..4149 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 4279..4299 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 4724..4858 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 4900..4933 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 5090..5806 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 5090..5803 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(5132..5153) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5393..5412) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5740..5761 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5821..5838) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5827..6051 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 6194..6315 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(6383..6401) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(6422..6438) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6422..6438) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6435..6457) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6446..6462 /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(6470..6500) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6515..6536 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6653..6670) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6824..7412) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6904..6923) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7583..8443) /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 8206..8225 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8444..8548) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct 481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg 661 gccccccctc gaggtcgacg gtatcgataa gcttccctta attaaccgtt taaacaattc 721 tgcaggaatc tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 781 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 841 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 901 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 961 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 1021 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 1081 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc 1141 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg 1201 cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg 1261 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta 1321 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcggggagtc 1381 gctgcgacgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg 1441 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg 1501 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct 1561 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg 1621 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg 1681 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg 1741 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt 1801 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc 1861 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc 1921 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc 1981 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg 2041 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg 2101 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc 2161 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct 2221 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg 2281 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg 2341 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg 2401 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgattt gataccgcgg 2461 gctcgacact agtgaacctc ttcgagggac ctaataactt cgtatagcat acattatacg 2521 aagttatatt aagggttccg gatccatggg ttgctgtttc tccaagacca tggtgagcaa 2581 gggagaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa 2641 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac 2701 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc 2761 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa 2821 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg 2881 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa 2941 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg 3001 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca 3061 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc 3121 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc 3181 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct 3241 gttcctgggg catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga 3301 ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc 3361 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac 3421 ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct 3481 gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga 3541 ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga 3601 cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa 3661 ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat 3721 gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat 3781 ccaccaggcc ctgaagctga aggacggcgg ccgctacctg gtggagttca agaccatcta 3841 catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat 3901 cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca 3961 ccacctgttc ctgtacggca tggacgagct gtacaagtaa ctcgagggat ctttgtgaag 4021 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct 4081 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt 4141 gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat 4201 gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac 4261 tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct 4321 tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt 4381 gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat 4441 tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact 4501 ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc 4561 tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat 4621 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4681 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4741 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4801 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg 4861 acatggtaag taagcttggg ctgcaggtcg agggacctaa taacttcgta tagcatacat 4921 tatacgaagt tatattaagg gttccggatc atcaccgcgg atgggttgct gtttctccaa 4981 gaccgcagcg aagggagaag ccgccgcgga gaggcctggg gaggcggctg tggcctcgtc 5041 gccttccaaa gcgaacggac aggagaatgg ccacgtgaag gtaggatccg tgagcaaggg 5101 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 5161 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 5221 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 5281 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt 5341 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 5401 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 5461 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 5521 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa 5581 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca 5641 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca 5701 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 5761 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgctcgag 5821 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 5881 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 5941 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 6001 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagc 6061 atagggatcc ccgaattcta gataactgat cataatcagc cataccacat ttgtagaggt 6121 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc 6181 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 6241 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 6301 catcaatgta tcttatttaa attgggcgcg cccaactagt tctagagcgg ccgccaccgc 6361 ggtggagctc cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat 6421 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6481 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 6541 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 6601 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6661 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6721 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6781 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6841 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6901 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6961 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 7021 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7081 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7141 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7201 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7261 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7321 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7381 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7441 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7501 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 7561 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 7621 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7681 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7741 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7801 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 7861 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 7921 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 7981 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 8041 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 8101 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 8161 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 8221 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8281 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8341 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8401 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8461 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 8521 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgc //