LOCUS V032696 8345 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032696 VERSION V032696 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 757..1980 /label="ZSCAN1(NM_182572)" /note="ZSCAN1(NM_182572)" /gene="ZSCAN1" CDS 2287..2343 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 2344..3054 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK*" /label="mCherry" misc_feature 3121..3709 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3775..3823 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4025..4453 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4467..4796 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4863..5657 /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 5833..5954 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6003..6019) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6027..6043 /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(6051..6081) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6096..6117 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6405..6993) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7164..8024) /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(8025..8129) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgtttaa acttaagctt gccaccatgc ttccacggcc caaagcccct 781 gcctccccca gacgccccca gaccccaacc ccgagtgagc aggacgcaga ccctgggcca 841 gcaagcccca gggacaccga agcccagcgt ctgcgcttcc ggcagttcca gtaccacgtg 901 gcgagcgggc cgcacctcgc gctgggccag ctctggacgc tgtgccgcca gtggctgagg 961 cccgaggcgc gctccaagga gcagatgctg gagctgctgg tgctggagca gttcctgggc 1021 gcgctgccca gcaagatgcg gacctgggtg cagtcacagg gcccccgaag ctgcagggag 1081 gccgccagcc tggtggagga cctcacacag atgtgccagc aggaagttct ggtatctctg 1141 gactcggtcg aaccccagga ctggagtttc ggtgaggagg aagatgggaa gagtccaagg 1201 tcccagaaag aaccatcgca ggcgtctgag ctgattctgg atgcagtggc agcagcccca 1261 gcactccccg aggaaagtga gtggctggag actacccagc tccagcagag tctgcacacc 1321 agggcggagg ccgaagcgcc ccgcgcccct ggcttgctgg ggtcccgggc ccgcttgcct 1381 ctgaagccga gtatctggga cgagcctgag gaccttctcg cagggccctc ctcagacctg 1441 cgggcagaag ggactgtgat ctcgagcccc aagggtccaa gtgctcagag aatcagtccc 1501 cgaaggagaa acaggaacac tgaccagagc ggccgccacc agccatccct caagcacacc 1561 aaaggtggta cccaagaggc tgttgcaggc atctcggtag tgccgcgtgg gccccgaggt 1621 gggcggccct tccagtgtgc cgactgtggg atggtcttca cctgggtcac ccacttcatc 1681 gagcaccaga agacccatcg cgaggaaggg ccctttccgt gccccgagtg tggcaaggtc 1741 ttcctgcaca actccgtcct cactgagcat ggcaagatcc acctgctgga gccaccgagg 1801 aagaaagccc cccggagcaa gggcccccgg gagtccgtcc cacccaggga tggagcccag 1861 ggcccagtgg cccctcgcag ccccaaaaga cccttccagt gtagcgtctg cgggaaggcc 1921 ttcccctgga tggtccacct cattgaccac cagaagctcc acacggccca cggccacatg 1981 gaattcgcca ccatgattct ctgggaaaag aaaagcgaag atattaccga gacattcaac 2041 cccgacaaca acatgatcgc cgccgacgcg agcgaggcat cggaggcggg gaaggcagaa 2101 aacacccatt acaccggcaa gcaccgcttc atgtaccgga tcctgttcgt cctgacctgc 2161 ctgatcacct tcattgtggt ggctattgtc ggcggaatta tcgcttcgaa cagaaacgtc 2221 ggaaatagaa tagctcaagg ccaaggatta gccttgaaat tagcaggtct tgatatcgga 2281 agcggagcta ctaacttcag cctgctgaag caggctggcg acgtggagga gaaccctgga 2341 cctatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc 2401 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag 2461 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg 2521 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag 2581 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag 2641 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag 2701 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc 2761 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac 2821 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac 2881 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac 2941 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac 3001 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaactcgag 3061 tctagagggc cccttaaggg ttcgatccct accggttagt aatgagtttg atatctcgac 3121 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 3181 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 3241 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 3301 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 3361 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 3421 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 3481 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 3541 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 3601 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 3661 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 3721 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 3781 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 3841 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 3901 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 3961 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 4021 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 4081 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 4141 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 4201 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 4261 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 4321 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 4381 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 4441 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 4501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4621 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 4681 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 4741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 4801 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 4861 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4921 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4981 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 5041 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 5101 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 5161 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 5221 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 5281 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 5341 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 5401 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 5461 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 5521 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 5581 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 5641 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 5701 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 5761 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 5821 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 5881 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5941 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 6001 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6061 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 6121 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 6181 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 6241 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 6301 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 6361 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6421 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6481 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6541 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 6601 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6661 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6721 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6781 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6841 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6901 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6961 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 7021 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 7081 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 7141 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 7201 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 7261 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 7321 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 7381 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 7441 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 7501 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7561 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7621 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7681 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7741 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7801 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7861 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7921 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7981 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 8041 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 8101 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 8161 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 8221 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 8281 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 8341 ttagg //