LOCUS V036325 8285 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036325 VERSION V036325 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 983..1006 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 1007..2659 /label="Gpr107(NM_178760)" /note="Gpr107(NM_178760)" /gene="Gpr107" CDS 2759..2779 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2807..3808 /codon_start=1 /note="In the Tet-Off® system, tTA binds to the Tet-responsive element and stimulates transcription only in the absence of tetracycline or doxycycline." /product="tetracycline-controlled transactivator, comprising a fusion of the tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16" /transl_table=1 /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS,AYSRARTKNNYGSTIE GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG*" /label="tTA" polyA_signal 3852..4076 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4122..4550 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4564..4893 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4960..5754 /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 5928..6049 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6098..6114) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6122..6138 /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(6146..6176) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6191..6212 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6500..7085) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7256..8116) /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(8117..8221) /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 aagacgatca tcgccctgag ctacatcttc tgcctggtat tcgccgacta 961 caaggacgat gatgacgcca gcgactacaa ggacgacgat gacaagatgg ccgtcccggt 1021 acccctcggc cgcttcggct ccttctgcct gcggctgcta cgactgctgg cgttgctgga 1081 gctgctggtc caccccgtct tgggccgcgt ccatcacctg gcgctcaaag atgatgtgcg 1141 gcataaagtc catctcaaca ccttcgggtt cttcaaggat gggtacatgg tggtcaacgt 1201 cagcagcctc tctgtgaacg agcctgaggg agccacggac aaggatgctg agatcggatt 1261 cagtctggat cggactaaga acgatggctt ttcttcttac ctggatgaag atgtgaatta 1321 ctgtatttta aaaaagaagt ccatgtcttc tgtcactctt gtcatcttag acatctctgg 1381 aagcatagtc aaggtcagat ccccaccaga agctggtaag cagttaccag agattgtctt 1441 cagcaaggat gagaaaatcc tgagtcagag ccaggagcct gctgtcagct ccaaccccaa 1501 agacagtgag gctcggagaa ccctcgatgg tttcaaggct ggacgaagta cagtggactc 1561 aaaggctatc acagagcgat ccttctcaat tcacaagaat gacggggtgg tctcatttca 1621 gttcttcttt aacatcagca ccgatgacca ggaaggcctc tacagccttt acttccacaa 1681 gtgctcaggc aacaacgtga agcctggtga acaggcctca ttcagcctga atattgctat 1741 cacagagaag aaccctaaca gctacctctc tgccggggaa atccctctcc ccaagttata 1801 tgtttccatg gccttgttct tctttctgtc tgggaccatt tggattcaca tccttcgtaa 1861 gagaaggaat gatgtattta aaattcactg gttgatggcc gcccttcctt tcaccaagtc 1921 tctctcgttg gtgttccatg caatcgacta ccactacatc tcctcgcagg gctttccgat 1981 tgaaggctgg gctgttgtat actacataac tcacctgcta aagggcgcac tcctgttcat 2041 caccatcgcc ctcatcggca ctggctgggc cttcatcaag cacatcctgt ctgataaaga 2101 caagaagatc ttcatgattg tcatcccgct ccaggtcctg gcgaatgtgg cctacatcat 2161 catagagtcc actgaggagg gcacaactga gtatggccta tggaaggact ctctgttcct 2221 ggttgacttg ctttgctgtg gcgccatcct cttcccagtg gtgtggtcaa tcaggcattt 2281 acaagaagcc tccgccacag atggaaaagc tgccattaac ttagcgaagc tgaggctttt 2341 cagacattac tacgtcttga ttgtgtgcta catctacttt accaggatca tcgccttcct 2401 tctcaagttt gctgttccat tccagtggaa gtggctctac cagctgctgg atgaaacagc 2461 cacgctggtg ttctttgtcc tgacggggta taaatttcgc ccagcttcag ataaccccta 2521 cctgcagctc tctcaggaag atgatgacct ggagatggag tccgtagtga cgacatcggg 2581 tgtgatggag aatatgaaga aggtgaagaa agtgagcaac ggtgctgtgg agccccaggg 2641 cagctgggaa ggcaccgcgg atatcatcga taccggtgga cgcaccccac ccagcctggg 2701 tccccaagat gagtcctgca ccaccgccag ctcctccctg gccaaggaca cttcatcgga 2761 aaacctgtac ttccaatcca ccggtgagaa cctgtacttc cagctaagat tagataaaag 2821 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac 2881 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa 2941 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt 3001 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag 3061 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga 3121 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact 3181 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga 3241 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc 3301 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt 3361 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc 3421 cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga 3481 tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct 3541 ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga 3601 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt 3661 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc 3721 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc 3781 ccttggaatt gacgagtacg gtgggtagct cgagtctaga gggcccgttt aaacccgctg 3841 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 3901 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 3961 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 4021 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 4081 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 4141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 4201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 4261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 4321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 4381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 4441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 4501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 4561 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 4621 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 4681 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 4741 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 4801 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 4861 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 4921 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 4981 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 5041 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 5101 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 5161 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 5221 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 5281 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 5341 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 5401 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 5461 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 5521 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 5581 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 5641 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 5701 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 5761 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 5821 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 5881 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 5941 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 6001 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 6061 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 6121 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 6181 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 6241 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 6301 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 6361 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 6421 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6601 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 6661 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 6721 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 6841 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 6901 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 6961 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 7021 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 7081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 7141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 7201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 7261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 7321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 7381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 7441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 7501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 7561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 7861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 7921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 7981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 8041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 8101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 8161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 8221 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct 8281 atggt //