LOCUS Exported 4731 bp ds-DNA circular SYN 16-AUG-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS mVenus C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4731) AUTHORS Koushik SV, Chen H, Thaler C, Puhl HL, Vogel SS TITLE Cerulean, Venus, and VenusY67C FRET reference standards. JOURNAL Biophys J. 2006 Dec 15. 91(12):L99-L101. PUBMED 17040988 REFERENCE 2 (bases 1 to 4731) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4731 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 83..386 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 387..590 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 541..561 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 629..638 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 635..1351 /codon_start=1 /product="Venus YFP with monomerizing A206K mutation (Nagai et al., 2002; Kremers et al., 2006)" /label=mVenus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(680..701) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(941..960) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1288..1309 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1352..1417 /label=MCS /note="multiple cloning site" polyA_signal 1541..1662 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1578..1597) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1632..1651 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1669..2124) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1806..1827) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2018..2037 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2151..2255 /gene="bla" /label=AmpR promoter promoter 2257..2614 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2280..2300) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2465..2600 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2527..2546 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2649..3443 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2703..2722) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3313..3332 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3631..3650) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3675..3722 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4051..4639 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4540..4559 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc 61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga 601 accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt 661 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 721 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctcatctg 781 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctcg gctacggcct 841 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 901 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 961 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 1021 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 1081 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg 1141 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat 1201 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag 1261 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 1321 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc 1381 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat 1441 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 1501 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 1561 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 1621 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg 1681 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 1741 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 1801 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 1861 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 1921 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 1981 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 2041 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 2101 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 2161 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 2221 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg 2281 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 2341 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 2401 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 2461 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 2521 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 2581 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 2641 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 2701 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 2761 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 2821 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 2881 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 2941 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3001 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3061 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3121 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 3181 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3241 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3301 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3361 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 3421 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 3481 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 3541 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 3601 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 3661 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 3721 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 3781 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 3841 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 3901 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 3961 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 4021 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 4081 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 4141 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 4201 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 4261 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 4321 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 4381 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 4441 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 4501 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 4561 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 4621 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 4681 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg a //