LOCUS       V027343                 7185 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027343
VERSION     V027343
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..1865
                     /label="HAO1(NM_017545)"
                     /note="HAO1(NM_017545)"
                     /gene="HAO1"
     CDS             1872..1937
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1961..2549
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2615..2663
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2865..3293
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3307..3636
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3703..4497
                     /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    4673..4794
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4843..4859)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4867..4883
                     /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(4891..4921)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4936..4957
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5245..5833)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6004..6864)
                     /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(6865..6969)
                     /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 ccaccatgct cccccggcta atttgtatca
      781 atgattatga acaacatgct aaatcagtac ttccaaagtc tatatatgac tattacaggt
      841 ctggggcaaa tgatgaagaa actttggctg ataatattgc agcattttcc agatggaagc
      901 tgtatccaag gatgctccgg aatgttgctg aaacagatct gtcgacttct gttttaggac
      961 agagggtcag catgccaata tgtgtggggg ctacggccat gcagcgcatg gctcatgtgg
     1021 acggcgagct tgccactgtg agagcctgtc agtccctggg aacgggcatg atgttgagtt
     1081 cctgggccac ctcctcaatt gaagaagtgg cggaagctgg tcctgaggca cttcgttggc
     1141 tgcaactgta tatctacaag gaccgagaag tcaccaagaa gctagtgcgg caggcagaga
     1201 agatgggcta caaggccata tttgtgacag tggacacacc ttacctgggc aaccgtctgg
     1261 atgatgtgcg taacagattc aaactgccgc cacaactcag gatgaaaaat tttgaaacca
     1321 gtactttatc attttctcct gaggaaaatt ttggagacga cagtggactt gctgcatatg
     1381 tggctaaagc aatagaccca tctatcagct gggaagatat caaatggctg agaagactga
     1441 catcattgcc aattgttgca aagggcattt tgagaggtga tgatgccagg gaggctgtta
     1501 aacatggctt gaatgggatc ttggtgtcga atcatggggc tcgacaactc gatggggtgc
     1561 cagccactat tgatgttctg ccagaaattg tggaggctgt ggaagggaag gtggaagtct
     1621 tcctggacgg gggtgtgcgg aaaggcactg atgttctgaa agctctggct cttggcgcca
     1681 aggctgtgtt tgtggggaga ccaatcgttt ggggcttagc tttccagggg gagaaaggtg
     1741 ttcaagatgt cctcgagata ctaaaggaag aattccggtt ggccatggct ctgagtgggt
     1801 gccagaatgt gaaagtcatc gacaagacat tggtgaggaa aaatcctttg gccgtttcca
     1861 agatcggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     1921 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     1981 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2041 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2101 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2161 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2221 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2281 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2341 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     2401 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2461 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2521 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     2581 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     2641 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     2701 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     2761 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     2821 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2881 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2941 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3001 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3061 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3121 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3181 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3241 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3301 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3481 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3541 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3601 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3661 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3721 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3781 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3841 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3901 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3961 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4021 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4081 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4141 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4201 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4261 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4321 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4381 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4441 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4501 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4561 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4621 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4681 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4741 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4801 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4861 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4921 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4981 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5041 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5101 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6961 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7021 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7081 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7141 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
