LOCUS Exported 5673 bp ds-DNA circular SYN 16-JUN-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS ECFP-Annexin-12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5673) TITLE Davidson Cyan Fluorescent Proteins REFERENCE 2 (bases 1 to 5673) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5673 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 547..556 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 553..1269 /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(598..619) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(859..878) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1206..1227 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2401..2522 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2438..2457) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2492..2511 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2529..2984) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2666..2687) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2878..2897 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3011..3115 /gene="bla" /label=AmpR promoter promoter 3117..3474 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3140..3160) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3325..3460 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3387..3406 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3509..4303 /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(3563..3582) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4173..4192 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4491..4510) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4535..4582 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4911..5499 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5400..5419 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 601 gtcgagctgg acggcgacgt aaacggccac aggttcagcg tgtccggcga gggcgagggc 661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 721 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc 781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 841 cgtaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 961 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac 1021 aagcagaaga acggcatcaa ggcccacttc aagatccgcc acaacatcga ggacggcagc 1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctatggc catggcaacc 1321 aaaggaggta ctgtcaaagc tgcttcagga ttcaatgcca tggaagatgc ccagaccctg 1381 aggaaggcca tgaaagggct cggcaccgat gaagacgcca ttattagcgt ccttgcctac 1441 cgcaacaccg cccagcgcca ggagatcagg acagcctaca agagcaccat cggcagggac 1501 ttgatagacg acctgaagtc agaactgagt ggcaacttcg agcaggtgat tgtggggatg 1561 atgacgccca cggtgctgta tgacgtgcaa gagctgcgaa gggccatgaa gggagccggc 1621 actgatgagg gctgcctaat tgagatcctg gcctcccgga cccctgagga gatccggcgc 1681 ataagccaaa cctaccagca gcaatatgga cggagccttg aagatgacat tcgctctgac 1741 acatcgttca tgttccagcg agtgctggtg tctctgtcag ctggtgggag ggatgaagga 1801 aattatctgg acgatgctct cgtgagacag gatgcccagg acctgtatga ggctggagag 1861 aagaaatggg ggacagatga ggtgaaattt ctaactgttc tctgttcccg gaaccgaaat 1921 cacctgttgc atgtgtttga tgaatacaaa aggatatcac agaaggatat tgaacagagt 1981 attaaatctg aaacatctgg tagctttgaa gatgctctgc tggctatagt aaagtgcatg 2041 aggaacaaat ctgcatattt tgctgaaaag ctctataaat cgatgaaggg cttgggcacc 2101 gatgataaca ccctcatcag agtgatggtt tctcgagcag aaattgacat gttggatatc 2161 cgggcacact tcaagagact ctatggaaag tctctgtact cgttcatcaa gggtgacaca 2221 tctggagact acaggaaagt actgcttgtt ctctgtggag gagatgatta aggatccacc 2281 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta 2341 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 2401 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 2461 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 2521 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 2581 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 2641 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 2701 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 2761 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 2821 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 2881 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 2941 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg 3001 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 3061 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga 3121 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 3181 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 3241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3301 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3361 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3421 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga 3481 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 3541 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 3601 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 3661 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc 3721 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 3781 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 3841 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 3901 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 3961 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 4021 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 4081 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 4141 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 4201 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 4261 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 4321 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4381 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4441 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg 4501 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg 4561 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 4621 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4681 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 4741 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt 4801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 4861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 4921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 4981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 5041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 5101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 5161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 5221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 5281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 5341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 5401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 5461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 5521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 5581 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg 5641 gtcattagtt catagcccat atatggagtt ccg //