LOCUS V024847 8162 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024847 VERSION V024847 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..3652 /label="DLG1(NM_001290983)" /note="DLG1(NM_001290983)" /gene="DLG1" CDS 3659..3724 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3762..3986 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4032..4460 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4474..4803 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4870..5664 /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 5838..5959 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6008..6024) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6032..6048 /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(6056..6086) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6101..6122 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6410..6995) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7166..8026) /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(8027..8131) /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 atgccggtcc ggaagcaaga 961 tacccagaga gcattgcacc ttttggagga atatcgttca aaactaagcc aaactgaaga 1021 cagacagctc agaagttcca tagaacgggt tattaacata tttcagagca acctctttca 1081 ggctttaata gatattcaag aattttatga agtgacctta ctggataatc caaaatgtat 1141 agatcgttca aagccgtctg aaccaattca acctgtgaat acttgggaga tttccagcct 1201 tccaagctct actgtgactt cagagacact gccaagcagc cttagcccta gtgtagagaa 1261 atacaggtat caggatgaag atacacctcc tcaagagcat atttccccac aaatcacaaa 1321 tgaagtgata ggtccagaat tggttcatgt ctcagagaag aacttatcag agattgagaa 1381 tgtccatgga tttgtttctc attctcatat ttcaccaata aagccaacag aagctgttct 1441 tccctctcct cccactgtcc ctgtgatccc tgtcctgcca gtccctgctg agaatactgt 1501 catcctaccc accataccac aggcaaatcc tcccccagta ctggtcaaca cagatagctt 1561 ggaaacacca acttacgtta atggcacaga tgcagattat gaatatgaag aaatcacact 1621 tgaaagggga aattcagggc ttggtttcag cattgcagga ggtacggaca acccacacat 1681 tggagatgac tcaagtattt tcattaccaa aattatcaca gggggagcag ccgcccaaga 1741 tggaagattg cgggtcaatg actgtatatt acgagtaaat gaagtagatg ttcgtgatgt 1801 aacacatagc aaagcagttg aagcgttgaa agaagcaggg tctattgtac gcttgtatgt 1861 aaaaagaagg aaaccagtgt cagaaaaaat aatggaaata aagctcatta aaggtcctaa 1921 aggtcttggg tttagcattg ctggaggtgt tggaaatcag catattcctg gggataatag 1981 catctatgta accaaaataa ttgaaggagg tgcagcacat aaggatggca aacttcagat 2041 tggagataaa cttttagcag tgaataacgt atgtttagaa gaagttactc atgaagaagc 2101 agtaactgcc ttaaagaaca catctgattt tgtttatttg aaagtggcaa aacccacaag 2161 tatgtatatg aatgatggct atgcaccacc tgatatcacc aactcttctt ctcagcctgt 2221 tgataaccat gttagcccat cttccttctt gggccagaca ccagcatctc cagccagata 2281 ctccccagtt tctaaagcag tacttggaga tgatgaaatt acaagggaac ctagaaaagt 2341 tgttcttcat cgtggctcaa cgggccttgg tttcaacatt gtaggaggag aagatggaga 2401 aggaatattt atttccttta tcttagccgg aggacctgct gatctaagtg gagagctcag 2461 aaaaggagat cgtattatat cggtaaacag tgttgacctc agagctgcta gtcatgagca 2521 ggcagcagct gcattgaaaa atgctggcca ggctgtcaca attgttgcac aatatcgacc 2581 tgaagaatac agtcgttttg aagctaaaat acatgattta cgggagcaga tgatgaatag 2641 tagtattagt tcagggtcag gttctcttcg aactagccag aagcgatccc tctatgtcag 2701 agcccttttt gattatgaca agactaaaga cagtgggctt cccagtcagg gactgaactt 2761 caaatttgga gatatcctcc atgttattaa tgcttctgat gatgaatggt ggcaagccag 2821 gcaggttaca ccagatggtg agagcgatga ggtcggagtg attcccagta aacgcagagt 2881 tgagaagaaa gaacgagccc gattaaaaac agtgaaattc aattctaaaa cgagagataa 2941 aggggagatc cctgacgaca tgggatcaaa aggcctgaag catgtaactt ctaatgccag 3001 cgatagtgaa agtagttacc gtggtcaaga agaatacgtc ttatcttatg aaccagtgaa 3061 tcaacaagaa gttaattata ctcgaccagt gatcatattg ggacctatga aagacaggat 3121 aaatgatgac ttgatctcag aatttcctga caaatttgga tcctgtgttc ctcatacaac 3181 tagaccaaaa cgagattatg aggtagatgg aagagattat cattttgtga cttcaagaga 3241 gcagatggaa aaagatatcc aggaacataa attcattgaa gctggccagt ataacaatca 3301 tctatatgga acaagtgttc agtctgtacg agaagtagca gaaaagggca aacactgtat 3361 ccttgatgtg tctggaaatg ccataaagag attacagatt gcacagcttt accctatctc 3421 catttttatt aaacccaaat ccatggaaaa tatcatggaa atgaataagc gtctaacaga 3481 agaacaagcc agaaaaacat ttgagagagc catgaaactg gaacaggagt ttactgaaca 3541 tttcacagct attgtacagg gggatacgct ggaagacatt tacaaccaag tgaaacagat 3601 catagaagaa caatctggtt cttacatctg ggttccggca aaagaaaagc tactcgagga 3661 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 3721 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 3781 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 3841 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 3901 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 3961 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 4021 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 4081 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 4141 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 4201 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 4261 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 4321 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 4381 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 4441 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 4741 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4801 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 4861 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 4921 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 4981 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 5041 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 5101 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 5161 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 5221 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 5281 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 5341 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 5401 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 5461 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 5521 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 5581 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 5641 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 5701 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 5761 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 5821 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5881 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5941 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 6001 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 6061 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 6121 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6181 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 6241 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 6301 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 6361 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6421 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6481 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 6541 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 6601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 6661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 6721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6841 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 6961 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 7021 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 7081 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 7141 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 7201 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 7261 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 7321 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 7381 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 7441 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 7501 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 7561 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 7621 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 7681 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 7741 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 7801 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 7861 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 7921 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 7981 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 8041 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 8101 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 8161 tc //