LOCUS V032796 6332 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032796 VERSION V032796 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..1771 /label="Pdcd1(NM_008798)" /note="Pdcd1(NM_008798)" /gene="Pdcd1" CDS 1778..1804 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1808..1834 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1838..1864 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" polyA_signal 1899..2123 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2169..2597 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2611..2940 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3007..3801 /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 3975..4096 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4145..4161) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4169..4185 /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(4193..4223) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4238..4259 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4547..5132) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5303..6163) /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(6164..6268) /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 tgggtccggc aggtaccctg gtcattcact tgggctgtgc tgcagttgag 961 ctggcaatca gggtggcttc tagaggtccc caatgggccc tggaggtccc tcaccttcta 1021 cccagcctgg ctcacagtgt cagagggagc aaatgccacc ttcacctgca gcttgtccaa 1081 ctggtcggag gatcttatgc tgaactggaa ccgcctgagt cccagcaacc agactgaaaa 1141 acaggccgcc ttctgtaatg gtttgagcca acccgtccag gatgcccgct tccagatcat 1201 acagctgccc aacaggcatg acttccacat gaacatcctt gacacacggc gcaatgacag 1261 tggcatctac ctctgtgggg ccatctccct gcaccccaag gcaaaaatcg aggagagccc 1321 tggagcagag ctcgtggtaa cagagagaat cctggagacc tcaacaagat atcccagccc 1381 ctcgcccaaa ccagaaggcc ggtttcaagg catggtcatt ggtatcatga gtgccctagt 1441 gggtatccct gtattgctgc tgctggcctg ggccctagct gtcttctgct caacaagtat 1501 gtcagaggcc agaggagctg gaagcaagga cgacactctg aaggaggagc cttcagcagc 1561 acctgtccct agtgtggcct atgaggagct ggacttccag ggacgagaga agacaccaga 1621 gctccctacc gcctgtgtgc acacagaata tgccaccatt gtcttcactg aagggctggg 1681 tgcctcggcc atgggacgta ggggctcagc tgatggcctg cagggtcctc ggcctccaag 1741 acatgaggat ggacattgtt cttggcctct tctcgagtac ccatacgacg tacctgatta 1801 tgcaacttat ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta 1861 cgcatgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc 1921 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 1981 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 2041 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 2101 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg 2161 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 2221 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 2281 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 2341 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 2401 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 2461 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 2521 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 2581 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 2641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 2701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2761 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 2821 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct 2881 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 2941 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg 3001 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 3061 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 3121 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 3181 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 3241 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 3301 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 3361 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 3421 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 3481 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 3541 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 3601 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 3661 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 3721 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 3781 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 3841 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 3901 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 3961 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4021 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4081 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 4141 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4201 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4261 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 4321 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 4381 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 4441 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 4501 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4561 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 4621 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 4681 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4741 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4801 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4861 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 4921 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 4981 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5041 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc 5101 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 5161 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 5221 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5281 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 5341 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 5401 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 5461 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 5521 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 5581 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 5641 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 5701 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 5761 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 5821 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 5881 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 5941 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6001 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6061 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6121 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 6181 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 6241 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 6301 acggatcggg agatctcccg atcccctatg gt //