LOCUS V025960 7742 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025960 VERSION V025960 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 941..3232 /label="PIGR(NM_002644)" /note="PIGR(NM_002644)" /gene="PIGR" CDS 3239..3304 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3342..3566 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3612..4040 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4054..4383 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4450..5244 /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 5418..5539 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5588..5604) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5612..5628 /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(5636..5666) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5681..5702 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5990..6575) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6746..7606) /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(7607..7711) /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 atccgccacc atgctgctct tcgtgctcac 961 ctgcctgctg gcggtcttcc cagccatctc cacgaagagt cccatatttg gtcccgagga 1021 ggtgaatagt gtggaaggta actcagtgtc catcacgtgc tactacccac ccacctctgt 1081 caaccggcac acccggaagt actggtgccg gcagggagct agaggtggct gcataaccct 1141 catctcctcg gagggctacg tctccagcaa atatgcaggc agggctaacc tcaccaactt 1201 cccggagaac ggcacatttg tggtgaacat tgcccagctg agccaggatg actccgggcg 1261 ctacaagtgt ggcctgggca tcaatagccg aggcctgtcc tttgatgtca gcctggaggt 1321 cagccagggt cctgggctcc taaatgacac taaagtctac acagtggacc tgggcagaac 1381 ggtgaccatc aactgccctt tcaagactga gaatgctcaa aagaggaagt ccttgtacaa 1441 gcagataggc ctgtaccctg tgctggtcat cgactccagt ggttatgtaa atcccaacta 1501 tacaggaaga atacgccttg atattcaggg tactggccag ttactgttca gcgttgtcat 1561 caaccaactc aggctcagcg atgctgggca gtatctctgc caggctgggg atgattccaa 1621 tagtaataag aagaatgctg acctccaagt gctaaagccc gagcccgagc tggtttatga 1681 agacctgagg ggctcagtga ccttccactg tgccctgggc cctgaggtgg caaacgtggc 1741 caaatttctg tgccgacaga gcagtgggga aaactgtgac gtggtcgtca acaccctggg 1801 gaagagggcc ccagcctttg agggcaggat cctgctcaac ccccaggaca aggatggctc 1861 attcagtgtg gtgatcacag gcctgaggaa ggaggatgca gggcgctacc tgtgtggagc 1921 ccattcggat ggtcagctgc aggaaggctc gcctatccag gcctggcaac tcttcgtcaa 1981 tgaggagtcc acgattcccc gcagccccac tgtggtgaag ggggtggcag gaggctctgt 2041 ggccgtgctc tgcccctaca accgtaagga aagcaaaagc atcaagtact ggtgtctctg 2101 ggaaggggcc cagaatggcc gctgccccct gctggtggac agcgaggggt gggttaaggc 2161 ccagtacgag ggccgcctct ccctgctgga ggagccaggc aacggcacct tcactgtcat 2221 cctcaaccag ctcaccagcc gggacgccgg cttctactgg tgtctgacca acggcgatac 2281 tctctggagg accaccgtgg agatcaagat tatcgaagga gaaccaaacc tcaaggtacc 2341 agggaatgtc acggctgtgc tgggagagac tctcaaggtc ccctgtcact ttccatgcaa 2401 attctcctcg tacgagaaat actggtgcaa gtggaataac acgggctgcc aggccctgcc 2461 cagccaagac gaaggcccca gcaaggcctt cgtgaactgt gacgagaaca gccggcttgt 2521 ctccctgacc ctgaacctgg tgaccagggc tgatgagggc tggtactggt gtggagtgaa 2581 gcagggccac ttctatggag agactgcagc cgtctatgtg gcagttgaag agaggaaggc 2641 agcggggtcc cgcgatgtca gcctagcgaa ggcagacgct gctcctgatg agaaggtgct 2701 agactctggt tttcgggaga ttgagaacaa agccattcag gatcccaggc tttttgcaga 2761 ggaaaaggcg gtggcagata caagagatca agccgatggg agcagagcat ctgtggattc 2821 cggcagctct gaggaacaag gtggaagctc cagagcgctg gtctccaccc tggtgcccct 2881 gggcctggtg ctggcagtgg gagccgtggc tgtgggggtg gccagagccc ggcacaggaa 2941 gaacgtcgac cgagtttcaa tcagaagcta caggacagac attagcatgt cagacttcga 3001 gaactccagg gaatttggag ccaatgacaa catgggagcc tcttcgatca ctcaggagac 3061 atccctcgga ggaaaagaag agtttgttgc caccactgag agcaccacag agaccaaaga 3121 acccaagaag gcaaaaaggt catccaagga ggaagccgag atggcctaca aagacttcct 3181 gctccagtcc agcaccgtgg ccgccgaggc ccaggacggc ccccaggaag ccctcgagga 3241 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 3301 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 3361 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 3421 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 3481 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 3541 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 3601 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 3661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 3721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 3781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 3841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 3901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 3961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 4021 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4201 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 4261 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 4321 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4381 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 4441 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 4501 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 4561 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 4621 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 4681 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 4741 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 4801 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 4861 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 4921 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 4981 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 5041 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 5101 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 5161 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 5221 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 5281 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 5341 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 5401 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5461 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5521 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5581 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5641 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5701 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 6541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 6601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 6661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 6721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 6781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 6841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 6901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 6961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 7021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 7081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 7141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 7201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 7261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 7321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 7381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 7441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 7501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 7561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 7621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 7681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 7741 tc //