LOCUS V035077 6904 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035077 VERSION V035077 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..2394 /label="MMP3(NM_002422)" /note="MMP3(NM_002422)" /gene="MMP3" CDS 2401..2466 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2504..2728 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2774..3202 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3216..3545 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3612..4406 /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 4580..4701 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4750..4766) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4774..4790 /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(4798..4828) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4843..4864 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5152..5737) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5908..6768) /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(6769..6873) /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 accatgaaga gtcttccaat cctactgttg ctgtgcgtgg cagtttgctc agcctatcca 1021 ttggatggag ctgcaagggg tgaggacacc agcatgaacc ttgttcagaa atatctagaa 1081 aactactacg acctcaaaaa agatgtgaaa cagtttgtta ggagaaagga cagtggtcct 1141 gttgttaaaa aaatccgaga aatgcagaag ttccttggat tggaggtgac ggggaagctg 1201 gactccgaca ctctggaggt gatgcgcaag cccaggtgtg gagttcctga tgttggtcac 1261 ttcagaacct ttcctggcat cccgaagtgg aggaaaaccc accttacata caggattgtg 1321 aattatacac cagatttgcc aaaagatgct gttgattctg ctgttgagaa agctctgaaa 1381 gtctgggaag aggtgactcc actcacattc tccaggctgt atgaaggaga ggctgatata 1441 atgatctctt ttgcagttag agaacatgga gacttttacc cttttgatgg acctggaaat 1501 gttttggccc atgcctatgc ccctgggcca gggattaatg gagatgccca ctttgatgat 1561 gatgaacaat ggacaaagga tacaacaggg accaatttat ttctcgttgc tgctcatgaa 1621 attggccact ccctgggtct ctttcactca gccaacactg aagctttgat gtacccactc 1681 tatcactcac tcacagacct gactcggttc cgcctgtctc aagatgatat aaatggcatt 1741 cagtccctct atggacctcc ccctgactcc cctgagaccc ccctggtacc cacggaacct 1801 gtccctccag aacctgggac gccagccaac tgtgatcctg ctttgtcctt tgatgctgtc 1861 agcactctga ggggagaaat cctgatcttt aaagacaggc acttttggcg caaatccctc 1921 aggaagcttg aacctgaatt gcatttgatc tcttcatttt ggccatctct tccttcaggc 1981 gtggatgccg catatgaagt tactagcaag gacctcgttt tcatttttaa aggaaatcaa 2041 ttctgggcta tcagaggaaa tgaggtacga gctggatacc caagaggcat ccacacccta 2101 ggtttccctc caaccgtgag gaaaatcgat gcagccattt ctgataagga aaagaacaaa 2161 acatatttct ttgtagagga caaatactgg agatttgatg agaagagaaa ttccatggag 2221 ccaggctttc ccaagcaaat agctgaagac tttccaggga ttgactcaaa gattgatgct 2281 gtttttgaag aatttgggtt cttttatttc tttactggat cttcacagtt ggagtttgac 2341 ccaaatgcaa agaaagtgac acacactttg aagagtaaca gctggcttaa ttgtctcgag 2401 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 2461 gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 2521 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 2581 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 2641 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2701 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2761 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 3181 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3241 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3361 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3421 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3481 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3541 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3601 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3661 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3721 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3781 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3841 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3901 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3961 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4021 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4081 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4141 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4201 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4261 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4321 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4381 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 4441 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4501 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4561 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4621 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4681 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4741 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4801 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4861 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4921 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4981 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5041 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 5461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 5521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5581 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 5701 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 5761 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 5821 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5881 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5941 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 6001 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 6061 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 6121 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 6181 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 6241 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 6301 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 6361 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 6421 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 6481 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 6541 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6601 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 6661 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 6721 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 6781 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6841 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6901 cgtc //