LOCUS V038069 6139 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038069 VERSION V038069 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..1629 /label="CLTRN(NM_020665)" /note="CLTRN(NM_020665)" /gene="CLTRN" CDS 1636..1701 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1739..1963 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2009..2437 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2451..2780 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2847..3641 /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 3815..3936 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3985..4001) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4009..4025 /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(4033..4063) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4078..4099 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4387..4972) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5143..6003) /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(6004..6108) /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 accatgttgt ggctgctctt ttttctggtg actgccattc atgctgaact ctgtcaacca 1021 ggtgcagaaa atgcttttaa agtgagactt agtatcagaa cagctctggg agataaagca 1081 tatgcctggg ataccaatga agaatacctc ttcaaagcga tggtagcttt ctccatgaga 1141 aaagttccca acagagaagc aacagaaatt tcccatgtcc tactttgcaa tgtaacccag 1201 agggtatcat tctggtttgt ggttacagac ccttcaaaaa atcacaccct tcctgctgtt 1261 gaggtgcaat cagccataag aatgaacaag aaccggatca acaatgcctt ctttctaaat 1321 gaccaaactc tggaattttt aaaaatccct tccacacttg caccacccat ggacccatct 1381 gtgcccatct ggattattat atttggtgtg atattttgca tcatcatagt tgcaattgca 1441 ctactgattt tatcagggat ctggcaacgt agaagaaaga acaaagaacc atctgaagtg 1501 gatgacgctg aagataagtg tgaaaacatg atcacaattg aaaatggcat cccctctgat 1561 cccctggaca tgaagggagg gcatattaat gatgccttca tgacagagga tgagaggctc 1621 acccctctcc tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac 1681 tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc agcctcgact 1741 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 1801 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 1861 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 1921 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 1981 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2041 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2101 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2161 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 2221 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 2281 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 2341 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 2401 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 2461 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 2521 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2581 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 2641 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 2701 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 2761 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 2821 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 2881 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 2941 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3001 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 3061 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3121 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3181 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 3241 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 3301 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 3361 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 3421 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 3481 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 3541 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 3601 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 3661 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 3721 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 3781 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 3841 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 3901 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 3961 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 4021 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 4081 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 4141 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 4201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 4261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 4321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 4381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 4441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 4501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 4561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 4621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 4681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 4741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 4801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 4861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 4921 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 5701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 5761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 5821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 5881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 5941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6121 aaaagtgcca cctgacgtc //