LOCUS V022641 6536 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022641 VERSION V022641 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..2026 /label="ACKR3(NM_020311)" /note="ACKR3(NM_020311)" /gene="ACKR3" CDS 2033..2098 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2136..2360 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2406..2834 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2848..3177 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3244..4038 /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 4212..4333 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4382..4398) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4406..4422 /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(4430..4460) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4475..4496 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4784..5369) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5540..6400) /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(6401..6505) /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 atggatctgc atctcttcga 961 ctactcagag ccagggaact tctcggacat cagctggcca tgcaacagca gcgactgcat 1021 cgtggtggac acggtgatgt gtcccaacat gcccaacaaa agcgtcctgc tctacacgct 1081 ctccttcatt tacattttca tcttcgtcat cggcatgatt gccaactccg tggtggtctg 1141 ggtgaatatc caggccaaga ccacaggcta tgacacgcac tgctacatct tgaacctggc 1201 cattgccgac ctgtgggttg tcctcaccat cccagtctgg gtggtcagtc tcgtgcagca 1261 caaccagtgg cccatgggcg agctcacgtg caaagtcaca cacctcatct tctccatcaa 1321 cctcttcggc agcattttct tcctcacgtg catgagcgtg gaccgctacc tctccatcac 1381 ctacttcacc aacaccccca gcagcaggaa gaagatggta cgccgtgtcg tctgcatcct 1441 ggtgtggctg ctggccttct gcgtgtctct gcctgacacc tactacctga agaccgtcac 1501 gtctgcgtcc aacaatgaga cctactgccg gtccttctac cccgagcaca gcatcaagga 1561 gtggctgatc ggcatggagc tggtctccgt tgtcttgggc tttgccgttc ccttctccat 1621 tatcgctgtc ttctacttcc tgctggccag agccatctcg gcgtccagtg accaggagaa 1681 gcacagcagc cggaagatca tcttctccta cgtggtggtc ttccttgtct gctggctgcc 1741 ctaccacgtg gcggtgctgc tggacatctt ctccatcctg cactacatcc ctttcacctg 1801 ccggctggag cacgccctct tcacggccct gcatgtcaca cagtgcctgt cgctggtgca 1861 ctgctgcgtc aaccctgtcc tctacagctt catcaatcgc aactacaggt acgagctgat 1921 gaaggccttc atcttcaagt actcggccaa aacagggctc accaagctca tcgatgcctc 1981 cagagtctca gagacggagt actctgcctt ggagcagagc accaaactcg aggactacaa 2041 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2101 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2161 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2221 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2281 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2341 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 2401 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2701 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2761 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2821 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 2941 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3001 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3061 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3121 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3181 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3241 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3301 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3361 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3421 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3481 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3541 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3601 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3661 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3721 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3781 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3841 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3901 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 3961 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4021 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4081 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4141 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4201 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4261 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4321 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4381 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4441 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4501 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4561 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4621 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4681 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4741 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4801 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4861 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4921 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 4981 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5041 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5101 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5161 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5221 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5281 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 5341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //