LOCUS Exported 5667 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS mTagBFP2-Annexin-12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5667) AUTHORS Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV TITLE An enhanced monomeric blue fluorescent protein with the high chemical stability of the chromophore. JOURNAL PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8. PUBMED 22174863 REFERENCE 2 (bases 1 to 5667) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5667 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 85..388 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 389..592 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 543..563 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 631..640 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 637..1347 /codon_start=1 /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /label=mTagBFP2 /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" polyA_signal 2479..2600 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2516..2535) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2570..2589 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2607..3062) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2744..2765) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2956..2975 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3089..3193 /gene="bla" /label=AmpR promoter promoter 3195..3552 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3218..3238) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3403..3538 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3465..3484 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3587..4381 /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(3641..3660) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4251..4270 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4569..4588) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4613..4660 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4989..5577 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5478..5497 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt 61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt 601 gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgtctaaggg cgaagagctg 661 attaaggaga acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc 721 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag 781 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac 841 ggcagcaaga ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc 901 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct 961 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg 1021 aacttcacat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc 1081 gagacgctgt accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc 1141 gtgggcggga gccatctgat cgcaaacgcc aagaccacat atagatccaa gaaacccgct 1201 aagaacctca agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag 1261 gccaacaacg agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc 1321 cctagcaaac tggggcacaa gcttaattcc ggactcagat ctcgagctca agcttcgaat 1381 tctatggcca tggcaaccaa aggaggtact gtcaaagctg cttcaggatt caatgccatg 1441 gaagatgccc agaccctgag gaaggccatg aaagggctcg gcaccgatga agacgccatt 1501 attagcgtcc ttgcctaccg caacaccgcc cagcgccagg agatcaggac agcctacaag 1561 agcaccatcg gcagggactt gatagacgac ctgaagtcag aactgagtgg caacttcgag 1621 caggtgattg tggggatgat gacgcccacg gtgctgtatg acgtgcaaga gctgcgaagg 1681 gccatgaagg gagccggcac tgatgagggc tgcctaattg agatcctggc ctcccggacc 1741 cctgaggaga tccggcgcat aagccaaacc taccagcagc aatatggacg gagccttgaa 1801 gatgacattc gctctgacac atcgttcatg ttccagcgag tgctggtgtc tctgtcagct 1861 ggtgggaggg atgaaggaaa ttatctggac gatgctctcg tgagacagga tgcccaggac 1921 ctgtatgagg ctggagagaa gaaatggggg acagatgagg tgaaatttct aactgttctc 1981 tgttcccgga accgaaatca cctgttgcat gtgtttgatg aatacaaaag gatatcacag 2041 aaggatattg aacagagtat taaatctgaa acatctggta gctttgaaga tgctctgctg 2101 gctatagtaa agtgcatgag gaacaaatct gcatattttg ctgaaaagct ctataaatcg 2161 atgaagggct tgggcaccga tgataacacc ctcatcagag tgatggtttc tcgagcagaa 2221 attgacatgt tggatatccg ggcacacttc aagagactct atggaaagtc tctgtactcg 2281 ttcatcaagg gtgacacatc tggagactac aggaaagtac tgcttgttct ctgtggagga 2341 gatgattaag gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta 2401 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 2461 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 2521 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 2581 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc 2641 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 2701 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 2761 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 2821 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 2881 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 2941 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 3001 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 3061 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 3121 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 3181 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3481 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3541 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 3601 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 3661 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 3721 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 3781 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 3841 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 3901 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 3961 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 4021 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 4081 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 4141 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 4201 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 4261 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 4321 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 4381 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 4441 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 4501 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 4561 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 4621 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 4681 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 4741 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 4801 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 4861 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 4921 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 4981 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 5041 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 5101 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 5161 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 5221 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 5281 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 5341 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 5401 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5461 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5521 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 5581 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 5641 tttcctgcgt tatcccctga ttctgtg //