LOCUS V039057 6577 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039057 VERSION V039057 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 928..2013 /label="GPR6(NM_005284)" /note="GPR6(NM_005284)" /gene="GPR6" CDS 2020..2043 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2050..2139 /codon_start=1 /product="two Strep-Tag®II moieties connected by a linker for enhanced binding to Strep-Tactin®, an engineered form of streptavidin (Schmidt et al., 2013)" /transl_table=1 /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK" /label="Twin-Strep-tag" polyA_signal 2177..2401 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2447..2875 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2889..3218 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3285..4079 /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 4253..4374 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4423..4439) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4447..4463 /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(4471..4501) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4516..4537 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4825..5410) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5581..6441) /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(6442..6546) /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 aagctgaatt cgccaccatg aacgcgagcg ccgcctcgct caacgactcc 961 caggtggtgg tagtggcggc cgaaggagcg gcggcggcgg ccacagcagc aggggggccg 1021 gacacgggcg aatggggacc ccctgctgcg gcggctctag gagccggcgg cggagctaat 1081 gggtctctgg agctgtcctc gcagctgtcg gctgggccac cgggactcct gctgccagcg 1141 gtgaatccgt gggacgtgct cctgtgcgtg tcggggacag tgatcgctgg agaaaacgcg 1201 ctggtggtgg cgctcatcgc gtccactccg gcgctgcgca cgcccatgtt cgtgctggta 1261 ggcagcctgg ccaccgctga cctgttggcg ggctgtggcc tcatcttgca ctttgtgttc 1321 cagtacttgg tgccctcgga gactgtgagt ctgctcacgg tgggcttcct cgtggcctcc 1381 ttcgccgcct ctgtcagcag cctgctggcc attacggtgg accgctacct gtccctgtat 1441 aacgcgctca cctattactc gcgccggacc ctgttgggcg tgcacctcct gcttgccgcc 1501 acttggaccg tgtccctagg cctggggctg ctgcccgtgc tgggctggaa ctgcctggca 1561 gagcgcgccg cctgcagcgt ggtgcgcccg ctggcgcgca gccacgtggc tctgctctcc 1621 gccgccttct tcatggtctt cggcatcatg ctgcacctgt acgtgcgcat ctgccaggtg 1681 gtctggcgcc acgcgcacca gatcgcgctg cagcagcact gcctggcgcc accccatctc 1741 gctgccacca gaaagggtgt gggtacactg gctgtggtgc tgggcacttt cggcgccagc 1801 tggctgccct tcgccatcta ttgcgtggtg ggcagccatg aggacccggc ggtctacact 1861 tacgccaccc tgctgcccgc cacctacaac tccatgatca atcccatcat ctatgccttc 1921 cgcaaccagg agatccagcg cgccctgtgg ctcctgctct gtggctgttt ccagtccaaa 1981 gtgccctttc gttccaggtc tcccagcgag gtcctcgagg attacaagga tgacgacgat 2041 aagggatcca gcgcctggtc ccaccctcag tttgagaagg gcggaggctc tggcggcgga 2101 agcggaggat ctgcttggag ccacccccag ttcgaaaagt gatgagggcc cgtttaaacc 2161 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2221 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2281 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2341 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2401 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2461 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2521 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2581 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 2641 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 2701 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 2761 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 2821 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 2881 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 2941 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3061 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3121 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3181 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3241 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3301 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3361 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3421 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3481 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3541 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3601 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3661 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3721 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3781 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 3841 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 3901 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 3961 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4021 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4081 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4141 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4201 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4381 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4441 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4501 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4561 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4621 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4681 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 4741 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 4801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 4861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 4921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 4981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5341 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5521 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5581 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5641 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5701 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5761 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5821 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5881 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5941 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6001 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6061 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6121 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6181 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6241 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6301 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6361 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6421 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6541 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //