LOCUS V043035 7615 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043035 VERSION V043035 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 964..3105 /label="SLC12A8(NM_024628)" /note="SLC12A8(NM_024628)" /gene="SLC12A8" CDS 3112..3177 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3215..3439 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3485..3913 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3927..4256 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4323..5117 /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 5291..5412 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5461..5477) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5485..5501 /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(5509..5539) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5554..5575 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5863..6448) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6619..7479) /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(7480..7584) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgaccc agatgtccca ggtgcaggag ctcttccatg aggcagccca gcaggatgcc 1021 ctggcccagc cccagccctg gtggaagacc cagctgttca tgtgggagcc tgtgctgttt 1081 gggacctggg atggtgtgtt cacatcctgc atgatcaaca tctttggggt tgtgctcttc 1141 ctgaggactg gctggctggt gggaaacaca ggagtgctcc tgggcatgtt cctggtgtcc 1201 ttcgtcatcc tggtggccct cgtcacggtg ctgtctggca ttggcgtcgg ggagcgcagc 1261 agcatcggca gcggtggcgt ctactccatg atctcctcgg tcctgggtgg gcagacggga 1321 ggcaccatcg ggctgctcta tgtgtttgga cagtgtgttg caggtgccat gtatatcacc 1381 ggctttgctg aatccatctc ggatttgctg ggcctcggga atatctgggc tgtgcgagga 1441 atttcagttg cggtgcttct ggccttgctg ggcattaacc tcgcaggtgt caaatggata 1501 atccgcctcc agctgctgtt gctgttcctg ctggccgtgt ccacactgga ctttgtggtg 1561 ggttctttca cccacctgga cccagaacat ggtttcattg gatattcacc cgaactgcta 1621 cagaacaaca cgctgcccga ttacagcccg ggggaatctt ttttcactgt ctttggggtt 1681 ttcttcccag cggctacagg agtcatggcc ggcttcaaca tggggggcga cctcagggag 1741 cctgccgcca gcattcccct gggctccctg gcagctgttg gcatctcgtg gtttctgtac 1801 atcatcttcg tcttcctcct gggcgccatc tgcactcgag aggcccttcg ctatgacttc 1861 ctgatagcgg aaaaggtatc cctcatgggc ttcctgttcc ttttgggctt atacatctcg 1921 tccctggctt cctgcatggg aggactttat ggagctcccc gcatcctgca gtgcattgcc 1981 caggagaaag tgatccctgc acttgcctgt ctgggacaag ggaaggggcc aaacaaaaca 2041 cccgtggctg ccatctgcct gaccagcttg gtgaccatgg cctttgtttt tgtgggtcaa 2101 gtgaacgttc tggcccccat cgtcaccatc aacttcatgc tgacatacgt tgcagtggac 2161 tactcttact tctccctgtc catgtgttcc tgcagcctga ccccggtgcc tgagccggtg 2221 ctcagggagg gcgcagaagg cctccactgc tctgagcacc tgctcttaga gaaagctccc 2281 agttacggct ctgagggacc tgcccaaaga gtcttggagg gcacgctact ggaattcacc 2341 aaggacatgg atcagctcct ccagctaacc aggaagcttg agagtagcca gcccaggcaa 2401 ggagagggta acaggacccc agaaagtcag aagaggaaaa gcaagaaggc caccaagcag 2461 accctacaag atagcttcct cttggacctc aaatcccctc cttctttccc tgtcgagatc 2521 tctgacaggt tgcccgctgc ctcctgggag gggcaggagt cctgctggaa caagcagact 2581 tccaagagcg aagggactca gcctgaggga acatatggag agcaacttgt tcctgagctg 2641 tgcaaccaat cagagtccag tggagaagat ttcttcctga agtccaggct ccaagaacaa 2701 gatgtctgga gaagatccac ttctttctat acccacatgt gcaacccctg ggtctccctg 2761 ttgggggctg ttgggtccct tctcatcatg tttgtgatac agtgggtgta taccctggtt 2821 aacatgggtg ttgctgccat cgtgtatttc tacattggcc gggccagtcc agggcttcac 2881 cttggatcag cctccaactt cagctttttc cggtggatga ggtctctctt gctcccctcc 2941 tgcaggagct tgcggtcccc tcaggagcag atcatcttgg cgccgtccct ggctaaggtt 3001 gacatggaga tgactcagct cacccaggag aatgcagact tcgccactcg ggatcgctac 3061 caccactcct ccctcgtgaa ccgggagcag ctgatgcctc actacctcga ggactacaaa 3121 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3181 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 3241 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 3301 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 3361 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 3421 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 3481 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 3541 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 3601 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 3661 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 3721 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 3781 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3841 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 3901 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 4021 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4081 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 4141 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 4201 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 4261 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 4321 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 4501 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 4561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 4921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 5041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 5101 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 5161 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 5221 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 5281 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 5341 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 5401 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 5461 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 5521 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 5581 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 5641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 5701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 5761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 5821 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 5881 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5941 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6001 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6061 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6121 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6181 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6241 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6301 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6361 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 6421 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 6481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 6541 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 6601 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 6661 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 6721 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 6781 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 6841 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 6901 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6961 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 7021 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 7081 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 7141 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 7201 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 7261 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 7321 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 7381 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 7441 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 7501 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 7561 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //