LOCUS Exported 6546 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS XE122 XDsh delta DIX-GFP-CS2+ SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6546) TITLE Xenopus Expression Plasmids REFERENCE 2 (bases 1 to 6546) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6546 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 23..41 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 23..40 /label=SP6 /note="SP6 promoter, forward primer" CDS 1819..2535 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(1849..1877) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2472..2492 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" promoter complement(2564..2581) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2586..2720 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2610..2629) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2664..2683 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2833..2853) /label=T3 /note="T3 promoter, forward primer" promoter complement(2833..2851) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2872..2888) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2872..2888) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2885..2907) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2896..2912 /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(2920..2950) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2965..2986 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3103..3120) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3274..3862) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3354..3373) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4033..4893) /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 4656..4675 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4894..4998) /gene="bla" /label=AmpR promoter rep_origin complement(5024..5479) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5161..5182) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5373..5392 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5557..6534 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct acttgttctt 61 tttgcaggat ctaccatgag tgagcaagca ggtgttggca gaggggtgaa tggacggaca 121 gagcggcacc tatctgggta tgaatcttcc tcgactctat tgaccagtga gattgagacg 181 agtatctgtg attcggagga ggacgacacc atgagcaggt tcagcagttc cacggagcag 241 agcagtgcct cacgccttct caaacggcac cggcgccgcc gcaagcagag acccccgcgc 301 ctggaacgga cgtcctcatt cagcagcgtc accgactcca ccatgtccct aaacatcatc 361 actgtgacgc tgaatatgga gaagtacaat ttcctgggca tctccatcgt ggggcagagt 421 aacgagcggg gagacggggg catttacatt ggctctataa tgaagggcgg agctgtggca 481 gctgacggca gaattgaacc cggggacatg ttgctacagg tgaatgacat taactttgag 541 aatatgagca atgacgatgc agtccgtgtg ctcagggata ttgtccataa accggggccc 601 attgtcctga ccgtggccaa atgctgggac ccttcccctc aggggtactt taccctcccc 661 cgcaatgagc ccatccaccc tattgatccg gctgcctggg tctctcactc ggctgccctg 721 agcgggtcct tcccagtgta ccccgggagc gcctccatga gcagcatgac ctccagcacc 781 tctgtcactg agactgagct ttcccatgcg ctccctccgg tgtcactctt cagcctctcc 841 gtacacaccg atttggcttc tgttgtgaag gtcatggctt cccctgagtc cggcctcgag 901 gtccgagatc gcatgtggct caaaatcacc attcccaacg ccttcctggg ttctgatgtg 961 gtggactggc tctatcacca cgtggaggga ttccaggacc ggcgagaggc tcgcaagttt 1021 gccagtaacc tgctgaaggc cggcttcatt cgccacacag tgaacaagat cacattctcc 1081 gagcagtgtt actatatctt tggggatctg actggctgtg agaactatat ggccaacctc 1141 tctctgaatg acaacgatgg atccagcggc gcttccgacc aggacactct ggcccctctt 1201 cccctccctg gggcttcccc atggcctcta ctccccacct tctcttacca gtatcaagcc 1261 ccacacccgt acagcactca gccacctgcc taccatgagc tctcctcgta cagctacggc 1321 atgggcagtg cgggcagcca gcacagcgag ggcagcagaa gcagtggatc caaccgcagc 1381 gatgggggtc ggggcatgca gaaggatgac cgttccggag tagcgggagt gggtggggga 1441 gactcgaagt cgggatctgg tagtgagtct gagtattcca cccgcagcag cataaggaga 1501 gtaggtggtg gggaggctgg tccccccagt gagcgctcca cttcatcccg acttccacct 1561 caccatccac cctctgttca ctcctatgct gctcctggtg tccctctctc ctacaatcca 1621 atgatgctca tgatgatgcc gccccctcca ttacctccac ctggggtttg tcctcccaat 1681 tcttctgtac cacctggagc tccacctctt gtacgggatc ttgcgtctgt tcccccggag 1741 ctgactgcca gccgccagtc tttccatatg gccatgggca accctagcga gttctttgtg 1801 gatgtcatgc atcgattgat gagtaaagga gaagaacttt tcactggagt tgtcccaatt 1861 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa 1921 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct 1981 gttccatggc caacacttgt cactactttc acttatggtg ttcaatgctt ttcaagatac 2041 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag 2101 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt 2161 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga 2221 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca 2281 gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga 2341 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt 2401 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag 2461 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat 2521 gaactataca aataagaatt caaggcctct cgagcctcta gaactatagt gagtcgtatt 2581 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 2641 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 2701 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 2761 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc 2821 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct 2881 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat 2941 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3001 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 3061 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 3121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 3181 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 3241 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 3301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 3361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 3421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 3481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 3541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 3601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 3661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 3721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 4141 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 4201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 4261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 4321 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 4381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 4441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 4501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 4561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 4621 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 4681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4861 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 4981 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat 5041 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 5101 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 5161 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 5221 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 5281 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 5341 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 5401 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 5461 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 5521 tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg 5581 actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg 5641 actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg 5701 caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg 5761 gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg 5821 gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata 5881 tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg tattggccct 5941 atgccatata gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg 6001 gcggtcccat ataccatata tggggcttcc taataccgcc catagccact cccccattga 6061 cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg 6121 tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc 6181 attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat 6241 atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat 6301 caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat 6361 tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat 6421 ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg 6481 ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat 6541 tctgcc //