LOCUS       V030287                 8523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030287
VERSION     V030287
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..3249
                     /label="SLCO5A1(NM_030958)"
                     /note="SLCO5A1(NM_030958)"
                     /gene="SLCO5A1"
     CDS             3256..3279
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    3299..3887
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3953..4001
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4203..4631
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4645..4974
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5041..5835
                     /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    6011..6132
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6181..6197)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6205..6221
                     /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(6229..6259)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6274..6295
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6583..7171)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7342..8202)
                     /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(8203..8307)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga cgaaggcact
      721 ggactgcagc ccggggcggg agagcagctg gaggcgccgg ccactgcaga agctgtccaa
      781 gagaggtgcg agccggagac cctcaggtct aagagtttac cggtcctcag cagcgcctcc
      841 tgccggccaa gcctcagtcc cactagtgga gacgccaacc cggcctttgg ctgtgtggat
      901 tcttcgggcc accaggagtt gaagcaaggc ccgaacccgt tggcccccag tccctctgcc
      961 ccgtccactt cggcggggct cggggactgt aaccacaggg tggacctcag caaaaccttc
     1021 tcggtgtcct ccgccttggc catgctccag gagagaaggt gcctctacgt ggtcctcacg
     1081 gattcccgtt gcttcctggt gtgcatgtgc tttctgacct tcatccaggc gttaatggtc
     1141 tctgggtacc tgagcagcgt aattaccacc attgaaaggc gctacagtct gaagagttcc
     1201 gagtcggggc tgctggtcag ctgctttgac atcgggaacc tggtggtggt ggtgttcgtc
     1261 agctacttcg gcggccgggg tcggcggccc ctgtggctgg ccgtgggtgg actcctcatc
     1321 gccttcgggg cagccctctt cgccttacct cacttcatct cgccccccta ccagatccaa
     1381 gagttgaacg cctcggcccc caacgacggc ctgtgtcagg gtggcaactc caccgccact
     1441 ttggagcctc cggcctgtcc gaaggactcg ggaggaaata atcactgggt ctacgtggct
     1501 ttattcattt gcgcgcagat tctcattgga atgggctcca cacctattta taccctggga
     1561 ccaacctact tagatgacaa tgtcaagaaa gaaaactcct ccttgtacct agccatcatg
     1621 tatgtcatgg gagcacttgg ccctgcagtg ggatatttat taggtggact tcttattggt
     1681 ttttatgttg atcccagaaa tcctgttcac cttgaccaga atgaccctcg tttcattgga
     1741 aactggtgga gtggattcct cctttgtgcc attgcaatgt ttcttgtgat attcccaatg
     1801 tttactttcc caaaaaagct tccacctcga cacaagaaaa agaaaaagaa aaaattttct
     1861 gttgatgctg ttagtgatga cgatgttctg aaggagaaat caaacaacag tgaacaagcg
     1921 gacaaaaaag tttcttcgat gggatttgga aaggatgtca gagacctacc aagagcagct
     1981 gtcaggatct taagcaacat gacattcctt tttgtgagtt tgtcatacac agctgagagt
     2041 gccattgtaa ctgctttcat taccttcatt cccaagttca tcgagtcaca gtttggtatc
     2101 ccagcctcca atgccagcat ctacactggg gttattatcg tccccagtgc tggtgttggt
     2161 attgtcctcg gaggctacat tataaaaaaa ttgaaacttg gtgccagaga atctgcaaaa
     2221 ctagcaatga tctgcagtgg tgtgtcttta ctatgttttt caaccctatt tattgttgga
     2281 tgtgaaagca ttaatctagg gggcataaac atcccttata caacaggacc ttctctcacc
     2341 atgccccata ggaatctgac aggaagctgc aacgttaatt gtggttgtaa aatacacgag
     2401 tatgagccag tctgtggatc agatggaatt acatacttta acccttgtct ggctggctgt
     2461 gttaatagtg gtaatcttag cactgggata cggaattata cagaatgcac ctgtgtccaa
     2521 agtcgccaag tgatcactcc acccaccgtg ggacagcgaa gtcagctccg tgtggttatt
     2581 gtcaagactt atctcaatga gaacggctat gctgtgtctg ggaaatgtaa acggacctgc
     2641 aatactctta tcccattctt agtttttctt ttcatagtca ccttcatcac agcatgtgcc
     2701 caaccatcag ctatcatagt aacactcagg tccgtagaag atgaggagag accttttgca
     2761 ctgggaatgc agtttgtttt gttgcgaaca cttgcataca ttcctactcc aatctacttt
     2821 ggagcagtca ttgacaccac ctgcatgctc tggcaacagg aatgtggtgt gcagggttct
     2881 tgctgggagt acaacgtgac gtcgtttcgt tttgtgtatt ttggtttggc tgccggcctc
     2941 aaattcgttg ggtttatttt tatttttctg gcctggtact ccataaaata caaggaggat
     3001 ggactgcaga ggcggaggca gagagaattt cccctgagca ccgtgagtga gagagtggga
     3061 caccccgaca atgcccggac tagatcttgc ccagctttca gcacccaggg agaattccac
     3121 gaagagactg gcctgcaaaa agggatccag tgcgcagcac agacctaccc ggggcccttc
     3181 ccagaagcaa taagttcctc tgcggacccg gggctggaag agagccccgc tgccttggag
     3241 ccgccctccc tcgagcacca tcaccatcac catcaccatt agggcccgat atctcgacaa
     3301 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3361 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3421 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3481 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3541 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3601 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3661 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3721 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3781 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3841 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     3901 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3961 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4021 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4081 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4141 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     4201 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4261 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4321 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4381 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4441 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4501 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4561 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4621 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4741 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4801 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4861 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4921 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4981 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5041 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5101 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5161 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5221 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5281 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5341 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5401 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5461 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5521 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5581 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5641 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5701 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5761 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5821 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     5881 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5941 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6001 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6121 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6181 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6241 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     6781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7501 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7561 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7621 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7681 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7741 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7801 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7861 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7921 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7981 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8041 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8101 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8161 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8221 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8281 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8341 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     8401 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8461 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8521 agg
//
