LOCUS V045515 7727 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045515 VERSION V045515 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..1624 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced monomeric variant of CFP (Goedhart et al., 2010)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNG IKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="mTurquoise" CDS 1661..2509 /label="VDAC1(NM_003374)" /note="VDAC1(NM_003374)" /gene="VDAC1" CDS 2540..3250 /codon_start=1 /product="bright monomeric yellow-green fluorescent protein derived from LanYFP (Shaner et al., 2013)" /transl_table=1 /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ KAFTDVMGMDELYK*" /label="mNeonGreen" polyA_signal 3294..3518 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3564..3992 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4006..4335 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4402..5196 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5370..5491 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5540..5556) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5564..5580 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(5588..5618) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5633..5654 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5942..6527) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6698..7558) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(7559..7663) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga 961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc 1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg 1081 gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca 1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 1321 ggggcacaag ctggagtaca actacttcag cgacaacgtc tatatcaccg ccgacaagca 1381 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca 1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 1501 caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca 1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 1621 caagggtgga ggcggatcag gaggcggtgg atctctcgag atggctgtgc cacccacgta 1681 tgccgatctt ggcaaatctg ccagggatgt cttcaccaag ggctatggat ttggcttaat 1741 aaagcttgat ttgaaaacaa aatctgagaa tggattggaa tttacaagct caggctcagc 1801 caacactgag accaccaaag tgacgggcag tctggaaacc aagtacagat ggactgagta 1861 cggcctgacg tttacagaga aatggaatac cgacaataca ctaggcaccg agattactgt 1921 ggaagatcag cttgcacgtg gactgaagct gaccttcgat tcatccttct cacctaacac 1981 tgggaaaaaa aatgctaaaa tcaagacagg gtacaagcgg gagcacatta acctgggctg 2041 cgacatggat ttcgacattg ctgggccttc catccggggt gctctggtgc taggttacga 2101 gggctggctg gccggctacc agatgaattt tgagactgca aaatcccgag tgacccagag 2161 caactttgca gttggctaca agactgatga attccagctt cacactaatg tgaatgacgg 2221 gacagagttt ggcggctcca tttaccagaa agtgaacaag aagttggaga ccgctgtcaa 2281 tcttgcctgg acagcaggaa acagtaacac gcgcttcgga atagcagcca agtatcagat 2341 tgaccctgac gcctgcttct cggctaaagt gaacaactcc agcctgatag gtttaggata 2401 cactcagact ctaaagccag gtattaaact gacactgtca gctcttctgg atggcaagaa 2461 cgtcaatgct ggtggccaca agcttggtct aggactggaa tttcaagcag gtggaggtgg 2521 atctggaggt ggtggatcaa tggtgagcaa gggcgaggag gataacatgg cctctctccc 2581 agcgacacat gagttacaca tctttggctc catcaacggt gtggactttg acatggtggg 2641 tcagggcacc ggcaatccaa atgatggtta tgaggagtta aacctgaagt ccaccaaggg 2701 tgacctccag ttctccccct ggattctggt ccctcatatc gggtatggct tccatcagta 2761 cctgccctac cctgacggga tgtcgccttt ccaggccgcc atggtagatg gctccggata 2821 ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc cttactgtta actaccgcta 2881 cacctacgag ggaagccaca tcaaaggaga ggcccaggtg aaggggactg gtttccctgc 2941 tgacggtcct gtgatgacca actcgctgac cgctgcggac tggtgcaggt cgaagaagac 3001 ttaccccaac gacaaaacca tcatcagtac ctttaagtgg agttacacca ctggaaatgg 3061 caagcgctac cggagcactg cgcggaccac ctacaccttt gccaagccaa tggcggctaa 3121 ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc actccaagac 3181 cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca tggacgagct 3241 gtacaagtaa ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 3301 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 3361 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 3421 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 3481 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 3541 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 3601 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 3661 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 3721 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 3781 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3841 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3901 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3961 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4081 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4141 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 4201 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 4261 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 4321 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 4381 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 4441 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 4501 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 4561 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 4621 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 4681 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 4741 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 4801 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 4861 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 4921 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 4981 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 5041 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 5101 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 5161 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 5221 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 5281 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 5341 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 5401 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 5461 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 5521 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 5581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 5641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 5701 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 5761 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5821 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5881 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5941 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6001 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6061 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 6181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 6241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 6301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 6361 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 6421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 6481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 6541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 6601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 6661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 6721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 6781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 6841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 6901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 6961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 7021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 7081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 7141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 7201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 7261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 7321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 7381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 7441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 7501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 7561 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 7621 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 7681 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt //