LOCUS V040547 6643 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040547 VERSION V040547 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..2079 /label="ACKR2(NM_001296)" /note="ACKR2(NM_001296)" /gene="ACKR2" CDS 2086..2109 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2116..2205 /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 2243..2467 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2513..2941 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2955..3284 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3351..4145 /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 4319..4440 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4489..4505) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4513..4529 /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(4537..4567) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4582..4603 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4891..5476) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5647..6507) /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(6508..6612) /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 gccgccactg cctctccgca gccactcgcc 961 actgaggatg ccgattctga gaatagcagc ttctattact atgactacct ggatgaagtg 1021 gccttcatgc tctgcaggaa ggatgcagtg gtgtcctttg gcaaagtctt cctcccagtc 1081 ttctatagcc tgatttttgt gttgggcctc agcgggaacc tccttcttct catggtcttg 1141 ctccgttacg tgcctcgcag gcggatggtt gagatctatc tgctgaatct ggccatctcc 1201 aaccttctgt ttctggtgac actgcccttc tggggcatct ccgtggcctg gcattgggtc 1261 ttcgggagtt tcttgtgcaa gatggtgagc actctttata ctattaactt ttacagtggc 1321 atctttttca ttagctgcat gagcctggac aagtacctgg agatcgttca tgctcagccc 1381 taccacaggc tgaggacccg ggccaagagc ctgctccttg ctaccatagt atgggctgtg 1441 tccctggccg tctccatccc tgatatggtc tttgtacaga cacatgaaaa tcccaagggt 1501 gtgtggaact gccacgcaga tttcggcggg catgggacca tttggaagct cttcctccgc 1561 ttccagcaga acctcctagg gtttctcctt ccactccttg ccatgatctt cttctactcc 1621 cgtattggtt gtgtcttggt gaggctgagg cccgcaggcc agggccgggc tttaaaaata 1681 gctgcagcct tggtggtggc cttcttcgtg ctatggttcc catacaatct caccttgttt 1741 ctgcatacgc tgttggacct gcaagtattc gggaactgtg aggtcagcca gcatctagac 1801 tacgcactcc aggtaacaga gagcatcgcc ttccttcact gctgcttttc ccccatcctg 1861 tatgccttct ccagtcaccg cttccgccag tacctgaagg ctttcctggc tgccgtgctt 1921 ggatggcacc tggcacctgg cactgcccag gcctcattat ccagctgttc tgagagcagc 1981 atacttactg cccaagagga aatgactggc atgaatgacc ttggagagag gcagtctgag 2041 aactacccta acaaggagga tgtggggaat aaatcagccc tcgaggatta caaggatgac 2101 gacgataagg gatccagcgc ctggtcccac cctcagtttg agaagggcgg aggctctggc 2161 ggcggaagcg gaggatctgc ttggagccac ccccagttcg aaaagtgatg agggcccgtt 2221 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 2281 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 2341 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 2401 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 2461 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc 2521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 2641 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 2701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 2761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 2821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 2881 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 2941 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 3061 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 3121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 3181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 3241 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt 3301 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac 3361 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3421 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3481 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 3541 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 3601 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 3661 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 3721 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 3781 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 3841 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 3901 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 3961 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 4021 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 4081 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 4141 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 4201 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4261 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa 4321 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4381 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 4441 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt 4501 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 4561 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 4621 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4681 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 4741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 4801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 4861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 4981 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 5101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 5341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 5401 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag 5461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5581 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5641 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5821 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6061 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6301 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6481 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6541 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6601 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //