LOCUS V032357 8831 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032357 VERSION V032357 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..4450 /label="ITGA2(NM_002203)" /note="ITGA2(NM_002203)" /gene="ITGA2" CDS 4457..4546 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" polyA_signal 4593..4817 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4863..5291 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5305..5634 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5701..6300 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 6474..6595 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6644..6660) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6668..6684 /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(6692..6722) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6737..6758 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7046..7631) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7802..8662) /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(8663..8767) /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 gggccagaac ggacaggggc cgcgccgctg ccgctgctgc tggtgttagc 961 gctcagtcaa ggcattttaa attgttgttt ggcctacaat gttggtctcc cagaagcaaa 1021 aatattttcc ggtccttcaa gtgaacagtt tggctatgca gtgcagcagt ttataaatcc 1081 aaaaggcaac tggttactgg ttggttcacc ctggagtggc tttcctgaga accgaatggg 1141 agatgtgtat aaatgtcctg ttgacctatc cactgccaca tgtgaaaaac taaatttgca 1201 aacttcaaca agcattccaa atgttactga gatgaaaacc aacatgagcc tcggcttgat 1261 cctcaccagg aacatgggaa ctggaggttt tctcacatgt ggtcctctgt gggcacagca 1321 atgtgggaat cagtattaca caacgggtgt gtgttctgac atcagtcctg attttcagct 1381 ctcagccagc ttctcacctg caactcagcc ctgcccttcc ctcatagatg ttgtggttgt 1441 gtgtgatgaa tcaaatagta tttatccttg ggatgcagta aagaattttt tggaaaaatt 1501 tgtacaaggc ctggatatag gccccacaaa gacacaggtg gggttaattc agtatgccaa 1561 taatccaaga gttgtgttta acttgaacac atataaaacc aaagaagaaa tgattgtagc 1621 aacatcccag acatcccaat atggtgggga cctcacaaac acattcggag caattcaata 1681 tgcaagaaaa tatgcttatt cagcagcttc tggtgggcga cgaagtgcta cgaaagtaat 1741 ggtagttgta actgacggtg aatcacatga tggttcaatg ttgaaagctg tgattgatca 1801 atgcaaccat gacaatatac tgaggtttgg catagcagtt cttgggtact taaacagaaa 1861 cgcccttgat actaaaaatt taataaaaga aataaaagca atcgctagta ttccaacaga 1921 aagatacttt ttcaatgtgt ctgatgaagc agctctacta gaaaaggctg ggacattagg 1981 agaacaaatt ttcagcattg aaggtactgt tcaaggagga gacaactttc agatggaaat 2041 gtcacaagtg ggattcagtg cagattactc ttctcaaaat gatattctga tgctgggtgc 2101 agtgggagct tttggctgga gtgggaccat tgtccagaag acatctcatg gccatttgat 2161 ctttcctaaa caagcctttg accaaattct gcaggacaga aatcacagtt catatttagg 2221 ttactctgtg gctgcaattt ctactggaga aagcactcac tttgttgctg gtgctcctcg 2281 ggcaaattat accggccaga tagtgctata tagtgtgaat gagaatggca atatcacggt 2341 tattcaggct caccgaggtg accagattgg ctcctatttt ggtagtgtgc tgtgttcagt 2401 tgatgtggat aaagacacca ttacagacgt gctcttggta ggtgcaccaa tgtacatgag 2461 tgacctaaag aaagaggaag gaagagtcta cctgtttact atcaaagagg gcattttggg 2521 tcagcaccaa tttcttgaag gccccgaggg cattgaaaac actcgatttg gttcagcaat 2581 tgcagctctt tcagacatca acatggatgg ctttaatgat gtgattgttg gttcaccact 2641 agaaaatcag aattctggag ctgtatacat ttacaatggt catcagggca ctatccgcac 2701 aaagtattcc cagaaaatct tgggatccga tggagccttt aggagccatc tccagtactt 2761 tgggaggtcc ttggatggct atggagattt aaatggggat tccatcaccg atgtgtctat 2821 tggtgccttt ggacaagtgg ttcaactctg gtcacaaagt attgctgatg tagctataga 2881 agcttcattc acaccagaaa aaatcacttt ggtcaacaag aatgctcaga taattctcaa 2941 actctgcttc agtgcaaagt tcagacctac taagcaaaac aatcaagtgg ccattgtata 3001 taacatcaca cttgatgcag atggattttc atccagagta acctccaggg ggttatttaa 3061 agaaaacaat gaaaggtgcc tgcagaagaa tatggtagta aatcaagcac agagttgccc 3121 cgagcacatc atttatatac aggagccctc tgatgttgtc aactctttgg atttgcgtgt 3181 ggacatcagt ctggaaaacc ctggcactag ccctgccctt gaagcctatt ctgagactgc 3241 caaggtcttc agtattcctt tccacaaaga ctgtggtgag gacggacttt gcatttctga 3301 tctagtccta gatgtccgac aaataccagc tgctcaagaa caacccttta ttgtcagcaa 3361 ccaaaacaaa aggttaacat tttcagtaac gctgaaaaat aaaagggaaa gtgcatacaa 3421 cactggaatt gttgttgatt tttcagaaaa cttgtttttt gcatcattct ccctgccggt 3481 tgatgggaca gaagtaacat gccaggtggc tgcatctcag aagtctgttg cctgcgatgt 3541 aggctaccct gctttaaaga gagaacaaca ggtgactttt actattaact ttgacttcaa 3601 tcttcaaaac cttcagaatc aggcgtctct cagtttccaa gccttaagtg aaagccaaga 3661 agaaaacaag gctgataatt tggtcaacct caaaattcct ctcctgtatg atgctgaaat 3721 tcacttaaca agatctacca acataaattt ttatgaaatc tcttcggatg ggaatgttcc 3781 ttcaatcgtg cacagttttg aagatgttgg tccaaaattc atcttctccc tgaaggtaac 3841 aacaggaagt gttccagtaa gcatggcaac tgtaatcatc cacatccctc agtataccaa 3901 agaaaagaac ccactgatgt acctaactgg ggtgcaaaca gacaaggctg gtgacatcag 3961 ttgtaatgca gatatcaatc cactgaaaat aggacaaaca tcttcttctg tatctttcaa 4021 aagtgaaaat ttcaggcaca ccaaagaatt gaactgcaga actgcttcct gtagtaatgt 4081 tacctgctgg ttgaaagacg ttcacatgaa aggagaatac tttgttaatg tgactaccag 4141 aatttggaac gggactttcg catcatcaac gttccagaca gtacagctaa cggcagctgc 4201 agaaatcaac acctataacc ctgagatata tgtgattgaa gataacactg ttacgattcc 4261 cctgatgata atgaaacctg atgagaaagc cgaagtacca acaggagtta taataggaag 4321 tataattgct ggaatccttt tgctgttagc tctggttgca attttatgga agctcggctt 4381 cttcaaaaga aaatatgaaa agatgaccaa aaatccagat gagattgatg agaccacaga 4441 gctcagtagc gatatcgaac aaaaactaat atcagaggaa gatttggaac agaaactaat 4501 cagtgaagaa gatttagagc aaaagttaat ttccgaggag gacttatagc tcgagtctag 4561 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 4621 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 4681 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 4741 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 4801 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 4861 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4921 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4981 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 5041 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 5101 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 5161 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 5221 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 5281 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 5341 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 5401 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 5461 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 5521 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 5581 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 5641 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 5701 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta 5761 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac 5821 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5881 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5941 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 6001 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 6061 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 6121 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 6181 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 6241 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 6301 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 6361 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 6421 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 6481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 6541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 6601 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 6661 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 6721 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 6781 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 6841 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 6901 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 6961 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 7501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7561 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 7621 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 7681 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 7741 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 7801 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 7861 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 7921 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 7981 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 8041 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 8101 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 8161 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 8221 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 8281 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 8341 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 8401 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8461 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 8521 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 8581 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 8641 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 8701 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 8761 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga 8821 tcccctatgg t //