LOCUS V040062 6637 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040062 VERSION V040062 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..2073 /label="S1PR1(NM_001320730)" /note="S1PR1(NM_001320730)" /gene="S1PR1" CDS 2080..2103 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2110..2199 /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 2237..2461 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2507..2935 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2949..3278 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3345..4139 /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 4313..4434 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4483..4499) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4507..4523 /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(4531..4561) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4576..4597 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4885..5470) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5641..6501) /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(6502..6606) /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 gggcccacca gcgtcccgct ggtcaaggcc 961 caccgcagct cggtctctga ctacgtcaac tatgatatca tcgtccggca ttacaactac 1021 acgggaaagc tgaatatcag cgcggacaag gagaacagca ttaaactgac ctcggtggtg 1081 ttcattctca tctgctgctt tatcatcctg gagaacatct ttgtcttgct gaccatttgg 1141 aaaaccaaga aattccaccg acccatgtac tattttattg gcaatctggc cctctcagac 1201 ctgttggcag gagtagccta cacagctaac ctgctcttgt ctggggccac cacctacaag 1261 ctcactcccg cccagtggtt tctgcgggaa gggagtatgt ttgtggccct gtcagcctcc 1321 gtgttcagtc tcctcgccat cgccattgag cgctatatca caatgctgaa aatgaaactc 1381 cacaacggga gcaataactt ccgcctcttc ctgctaatca gcgcctgctg ggtcatctcc 1441 ctcatcctgg gtggcctgcc tatcatgggc tggaactgca tcagtgcgct gtccagctgc 1501 tccaccgtgc tgccgctcta ccacaagcac tatatcctct tctgcaccac ggtcttcact 1561 ctgcttctgc tctccatcgt cattctgtac tgcagaatct actccttggt caggactcgg 1621 agccgccgcc tgacgttccg caagaacatt tccaaggcca gccgcagctc tgagaagtcg 1681 ctggcgctgc tcaagaccgt aattatcgtc ctgagcgtct tcatcgcctg ctgggcaccg 1741 ctcttcatcc tgctcctgct ggatgtgggc tgcaaggtga agacctgtga catcctcttc 1801 agagcggagt acttcctggt gttagctgtg ctcaactccg gcaccaaccc catcatttac 1861 actctgacca acaaggagat gcgtcgggcc ttcatccgga tcatgtcctg ctgcaagtgc 1921 ccgagcggag actctgctgg caaattcaag cgacccatca tcgccggcat ggaattcagc 1981 cgcagcaaat cggacaattc ctcccacccc cagaaagacg aaggggacaa cccagagacc 2041 attatgtctt ctggaaacgt caactcttct tccctcgagg attacaagga tgacgacgat 2101 aagggatcca gcgcctggtc ccaccctcag tttgagaagg gcggaggctc tggcggcgga 2161 agcggaggat ctgcttggag ccacccccag ttcgaaaagt gatgagggcc cgtttaaacc 2221 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2281 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2341 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2401 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2461 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 2701 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 2761 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 2821 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 2881 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 2941 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3001 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3181 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3241 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3301 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3361 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3421 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3481 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3541 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3601 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3661 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3721 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3781 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3841 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 3901 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 3961 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4021 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4081 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4141 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4201 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4261 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4321 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4381 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4441 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4501 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4561 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4621 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4681 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4741 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 4801 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 4861 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 4921 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 4981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5041 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5341 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5401 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6601 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //