LOCUS       V045635                 7725 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045635
VERSION     V045635
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2405
                     /label="SOAT1(NM_003101)"
                     /note="SOAT1(NM_003101)"
                     /gene="SOAT1"
     CDS             2412..2477
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2501..3089
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3155..3203
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3405..3833
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3847..4176
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4243..5037
                     /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    5213..5334
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5383..5399)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5407..5423
                     /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(5431..5461)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5476..5497
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5785..6373)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6544..7404)
                     /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(7405..7509)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt gggtgaagag aagatgtctc
      781 taagaaaccg gctgtcaaag tccagggaaa atcctgagga agatgaagac cagagaaacc
      841 ctgcaaagga gtccctagag acacctagta atggtcgaat tgacataaaa cagttgatag
      901 caaagaagat aaagttgaca gcagaggcag aggaattgaa gccatttttt atgaaggaag
      961 ttggcagtca ctttgatgat tttgtgacca atctcattga aaagtcagca tcattagata
     1021 atggtgggtg cgctctcaca accttttctg ttcttgaagg agagaaaaac aaccatagag
     1081 cgaaggattt gagagcacct ccagaacaag gaaagatttt tattgcaagg cgctctctct
     1141 tagatgaact gcttgaagtg gaccacatca gaacaatata tcacatgttt attgccctcc
     1201 tcattctctt tatcctcagc acacttgtag tagattacat tgatgaagga aggctggtgc
     1261 ttgagttcag cctcctgtct tatgcttttg gcaaatttcc taccgttgtt tggacctggt
     1321 ggatcatgtt cctgtctaca ttttcagttc cctattttct gtttcaacat tgggccactg
     1381 gctatagcaa gagttctcat ccgctgatcc gttctctctt ccatggcttt cttttcatga
     1441 tcttccagat tggagttcta ggttttggac caacatatgt tgtgttagca tatacactgc
     1501 caccagcttc ccggttcatc attatattcg agcagattcg ttttgtaatg aaggcccact
     1561 catttgtcag agagaacgtg cctcgggtac taaattcagc taaggagaaa tcaagcactg
     1621 ttccaatacc tacagtcaac cagtatttgt acttcttatt tgctcctacc cttatctacc
     1681 gtgacagcta tcccaggaat cccactgtaa gatggggtta tgtcgctatg aagtttgcac
     1741 aggtctttgg ttgctttttc tatgtgtact acatctttga aaggctttgt gcccccttgt
     1801 ttcggaatat caaacaggag cccttcagcg ctcgtgttct ggtcctatgt gtatttaact
     1861 ccatcttgcc aggtgtgctg attctcttcc ttactttttt tgcctttttg cactgctggc
     1921 tcaatgcctt tgctgagatg ttacgctttg gtgacaggat gttctataag gattggtgga
     1981 actccacgtc atactccaac tattatagaa cctggaatgt ggtggtccat gactggctat
     2041 attactatgc ttacaaggac tttctctggt ttttctccaa gagattcaaa tctgctgcca
     2101 tgttagctgt ctttgctgta tctgctgtag tacacgaata tgccttggct gtttgcttga
     2161 gctttttcta tcccgtgctc ttcgtgctct tcatgttctt tggaatggct ttcaacttca
     2221 ttgtcaatga tagtcggaaa aagccgattt ggaatgttct gatgtggact tctcttttct
     2281 tgggcaatgg agtcttactc tgcttttatt ctcaagaatg gtatgcacgt cagcactgtc
     2341 ctctgaaaaa tcccacattt ttggattatg tccggccacg ttcctggact tgtcgttacg
     2401 tgtttggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     2461 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     2521 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2581 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2641 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2701 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2761 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2821 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2881 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     2941 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3001 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3061 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3121 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3181 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3241 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3301 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3361 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3421 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3481 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3541 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3601 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3661 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3721 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3781 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3841 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4021 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4081 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4141 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4201 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4261 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4321 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4381 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4441 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4501 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4561 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4621 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4681 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4741 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4801 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4861 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4921 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4981 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5041 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5101 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5161 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5221 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5281 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5341 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5401 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5461 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5521 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5581 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6661 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6841 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7141 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7441 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7501 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7561 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7621 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7681 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
