LOCUS V024621 6364 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024621 VERSION V024621 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 931..1800 /label="Cd274(NM_021893)" /note="Cd274(NM_021893)" /gene="Cd274" CDS 1807..1896 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" polyA_signal 1931..2155 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2201..2629 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2643..2972 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3039..3833 /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 4007..4128 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4177..4193) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4201..4217 /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(4225..4255) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4270..4291 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4579..5164) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5335..6195) /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(6196..6300) /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 cactagtcca gtgtggtgga attcgccacc atgaggatat ttgctggcat tatattcaca 961 gcctgctgtc acttgctacg ggcgtttact atcacggctc caaaggactt gtacgtggtg 1021 gagtatggca gcaacgtcac gatggagtgc agattccctg tagaacggga gctggacctg 1081 cttgcgttag tggtgtactg ggaaaaggaa gatgagcaag tgattcagtt tgtggcagga 1141 gaggaggacc ttaagcctca gcacagcaac ttcaggggga gagcctcgct gccaaaggac 1201 cagcttttga agggaaatgc tgcccttcag atcacagacg tcaagctgca ggacgcaggc 1261 gtttactgct gcataatcag ctacggtggt gcggactaca agcgaatcac gctgaaagtc 1321 aatgccccat accgcaaaat caaccagaga atttccgtgg atccagccac ttctgagcat 1381 gaactaatat gtcaggccga gggttatcca gaagctgagg taatctggac aaacagtgac 1441 caccaacccg tgagtgggaa gagaagtgtc accacttccc ggacagaggg gatgcttctc 1501 aatgtgacca gcagtctgag ggtcaacgcc acagcgaatg atgttttcta ctgtacgttt 1561 tggagatcac agccagggca aaaccacaca gcggagctga tcatcccaga actgcctgca 1621 acacatcctc cacagaacag gactcactgg gtgcttctgg gatccatcct gttgttcctc 1681 attgtagtgt ccacggtcct cctcttcttg agaaaacaag tgagaatgct agatgtggag 1741 aaatgtggcg ttgaagatac aagctcaaaa aaccgaaatg atacacaatt cgaggagacg 1801 ctcgaggaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa 1861 gatttagagc aaaagttaat ttccgaggag gacttatgag ggcccgttta aacccgctga 1921 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 1981 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 2041 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2101 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2161 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2221 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2281 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2341 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2401 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2461 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2521 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 2581 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 2641 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2701 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 2761 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 2821 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 2881 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 2941 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 3001 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 3061 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 3121 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 3181 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 3241 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 3301 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 3361 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 3421 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 3481 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 3541 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 3601 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 3661 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 3721 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 3781 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 3841 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 3901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 3961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 4021 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 4081 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 4141 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 4321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 5041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 5101 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 5161 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 5221 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 5281 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 5341 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 5401 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 5461 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 5521 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 5581 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 5641 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 5701 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 5761 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 5821 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 5881 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 5941 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 6001 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 6061 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 6121 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 6181 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 6241 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 6301 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta 6361 tggt //