LOCUS V040270 7654 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040270 VERSION V040270 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..3144 /label="PLOD1(NM_000302)" /note="PLOD1(NM_000302)" /gene="PLOD1" CDS 3151..3216 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3254..3478 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3524..3952 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3966..4295 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4362..5156 /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 5330..5451 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5500..5516) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5524..5540 /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(5548..5578) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5593..5614 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5902..6487) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6658..7518) /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(7519..7623) /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 accatgcggc ccctgctgct actggccctg ctgggctggc tgctgctggc cgaagcgaag 1021 ggcgacgcca agccggagga caacctttta gtcctcacgg tggccactaa ggagaccgag 1081 ggattccgtc gcttcaagcg ctcagctcag ttcttcaact acaagatcca ggcgcttggc 1141 ctaggggagg actggaatgt ggagaagggg acgtcggcag gtggagggca gaaggtccgg 1201 ctgctgaaga aagctctgga gaagcacgca gacaaggagg atctggtcat tctcttcgca 1261 gacagctatg acgtgctgtt tgcatcgggg ccccgggagc tcctgaagaa gttccggcag 1321 gccaggagcc aggtggtctt ctctgctgag gagctcatct acccagaccg caggctggag 1381 accaagtatc cggtggtgtc cgatggcaag aggttcctgg gctctggagg cttcatcggt 1441 tatgccccca acctcagcaa actggtggcc gagtgggagg gccaggacag cgacagcgat 1501 cagctgtttt acaccaagat cttcttggac ccggagaaga gggagcagat caatatcacc 1561 ctggaccacc gctgccgtat cttccagaac ctggatggag ccttggatga ggtcgtgctc 1621 aagtttgaaa tgggccatgt gagagcgagg aacctggcct atgacaccct cccggtcctg 1681 atccatggca acgggccaac caagctgcag ttgaactacc tgggcaacta catcccgcgc 1741 ttctggacct tcgaaacagg ctgcaccgtg tgtgacgaag gcttgcgcag cctcaagggc 1801 attggggatg aagctctgcc cacggtcctg gtcggcgtgt tcatcgaaca gcccacgccg 1861 tttgtgtccc tgttcttcca gcggctcctg cggctccact acccccagaa acacatgcga 1921 cttttcatcc acaaccacga gcagcaccac aaggctcagg tggaagagtt cctggcacag 1981 catggcagcg agtaccagtc tgtgaagctg gtgggccctg aggtgcggat ggcgaatgca 2041 gatgccagga acatgggcgc agacctgtgc cggcaggacc gcagctgcac ctactacttc 2101 agcgtggatg ctgacgtggc cctgaccgag cccaacagcc tgcggctgct gatccaacag 2161 aacaagaacg tcattgcccc gctgatgacc cggcatggga ggctgtggtc gaacttctgg 2221 ggggctctca gtgcagatgg ctactatgcc cgttccgagg actacgtgga cattgtgcag 2281 gggcggcgtg ttggtgtctg gaatgtgccc tatatttcaa acatctactt gatcaagggc 2341 agtgccctgc ggggtgagct gcagtcctca gatctcttcc accacagcaa gctggacccc 2401 gacatggcct tctgtgccaa catccggcag caggatgtgt tcatgttcct gaccaaccgg 2461 cacacccttg gccatctgct ctccctagac agctaccgca ccacccacct gcacaacgac 2521 ctctgggagg tgttcagcaa ccccgaggac tggaaggaga agtacatcca ccagaactac 2581 accaaagccc tggcagggaa gctggtggag acgccctgcc cggatgtcta ttggttcccc 2641 atcttcacgg aggtggcctg tgatgagctg gtggaggaga tggagcactt tggccagtgg 2701 tctctgggca acaacaagga caaccgcatc cagggtggct acgagaacgt gccgactatt 2761 gacatccaca tgaaccagat cggctttgag cgggagtggc acaaattcct gctggagtac 2821 attgcgccca tgacggagaa gctctacccc ggctactaca ccagggccca gtttgacctg 2881 gcctttgtcg tccgctacaa gcctgatgag cagccctcac tgatgccaca ccatgatgcc 2941 tccaccttca ccatcaacat cgccctgaac cgagtcgggg tggattacga gggcgggggc 3001 tgtcggttcc tgcgctacaa ctgttccatc cgagccccaa ggaagggctg gaccctcatg 3061 caccctggac gactcacgca ttaccatgag gggctcccca ccaccagggg cacccgctac 3121 atcgcagtct ccttcgtcga tcccctcgag gactacaaag accatgacgg tgattataaa 3181 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc 3241 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 3301 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 3361 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 3421 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 3481 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 3541 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3601 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3661 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 3721 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3781 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3841 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 3901 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 4021 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 4141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 4201 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 4261 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 4321 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 4381 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 4441 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4501 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 4561 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4621 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4681 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4741 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4801 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4861 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4921 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4981 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 5041 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 5101 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 5161 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 5221 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 5281 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 5341 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5401 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5461 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 5521 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 5581 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5641 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5701 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 6121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 6241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 6301 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 6361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 6421 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 6841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 7021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7621 gcgcacattt ccccgaaaag tgccacctga cgtc //