LOCUS V045402 5812 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045402 VERSION V045402 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..1302 /label="RTL8A(NM_001078172)" /note="RTL8A(NM_001078172)" /gene="RTL8A" CDS 1309..1374 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1412..1636 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1682..2110 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2124..2453 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2520..3314 /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 3488..3609 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3658..3674) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3682..3698 /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(3706..3736) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3751..3772 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4060..4645) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4816..5676) /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(5677..5781) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggacg gtcgggtgca gctgatgaag gccctcctgg ccgggcccct ccggcccgcg 1021 gcgcgtcgct ggaggaaccc gattcccttt cccgagacgt ttgacggaga taccgaccga 1081 ctcccggagt tcatcgtgca gacgagctcc tacatgttcg tggacgagaa cacgttctcc 1141 aacgacgccc tgaaggtgac gttcctcatc acccgcctca cggggccagc cctgcagtgg 1201 gtgatcccct acatcaggaa ggagagcccc ctgctcaatg attaccgggg cttcctggcc 1261 gagatgaagc gggtctttgg atgggaggag gacgaggact tcctcgagga ctacaaagac 1321 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga 1381 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 1441 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 1501 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 1561 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 1621 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc 1681 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 1741 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 1801 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 1861 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 1921 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 1981 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 2041 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 2101 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 2161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 2221 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 2281 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 2341 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 2401 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 2461 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 2521 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 2581 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 2641 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 2701 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 2761 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 2821 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 2881 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 2941 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 3001 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 3061 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 3121 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 3181 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 3241 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 3301 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 3361 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 3421 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 3481 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3541 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 3601 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 3661 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 3721 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 3781 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3841 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 3901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 3961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 4501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat 4621 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 4681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 4741 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 4801 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 4861 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 4921 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 4981 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 5041 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 5101 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 5161 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 5221 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 5281 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 5341 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 5401 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 5461 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 5521 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 5581 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 5641 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 5701 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 5761 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //