LOCUS       V045141                 8445 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045141
VERSION     V045141
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..3125
                     /label="Iqub(NM_001034130)"
                     /note="Iqub(NM_001034130)"
                     /gene="Iqub"
     CDS             3132..3197
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3221..3809
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3875..3923
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4125..4553
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4567..4896
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4963..5757
                     /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    5933..6054
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6103..6119)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6127..6143
                     /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(6151..6181)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6196..6217
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6505..7093)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7264..8124)
                     /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(8125..8229)
                     /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 ccaccatgtc cgatacggag gagaatgtcc
      781 ttgactcccc ggccagtggt gaagcggccg gcgaagtcat cagtattcct ggccctacag
      841 acgaagcaga agagtcagat atctcaccag agcaagaata cgaaatagag ggatttacca
      901 ccgagagtga ggagacagtg gagcactaca gtgatctcag tgtttcaacc cccaaatctt
      961 acgactccaa acccagggac gaggtcgtgt cccttggaag tgccacagac tctcatgagc
     1021 aagaatacag acttggagat cttggaggtg accagagatc agaagaggac gagatgtcat
     1081 ttctttataa aatcaaagct ataaaggaga acttgcagtc gtcagtgaaa gagtgtatag
     1141 ccacagtaaa aattgtactt attcctgcag gccaggaaat tgtaatgcct ttcagagttg
     1201 atgctccatt tagattcctc aaggagcact ttgcacactt attacgtatc ccccatcatg
     1261 tattgcaaat ggcctataca ggaatcagtg ttgggaataa tgagtccctg atacaatatg
     1321 gagtcaaacc acgggaaatc gttcaaatga atgtattttc ctcagaaccc gaccaatatc
     1381 cagtcagaag aatagagggc ttgagtgagg gctccaagat cataacggtc accattcaga
     1441 ccggcatcga acgctacgag gaggtagctg tggagatagt aaaatctgac ttccacaagc
     1501 catttcttgg tggatttaga cataaaataa caggactaga ataccacaat gctgggacac
     1561 aaactgtacc caagaagatt ccggaaaaag acaatctgtt ctgtagggac acccagacag
     1621 tttttcagaa aaagaagctg caacaaacta tgaataccac ctccactcag atgactaaaa
     1681 tcggtgtata cgtatcaaac atgactgata aactgctcac gccaggaagt tacttttcag
     1741 ctgcagagta ccacgctcga aggttgaacg cggtgatagt gattcagact gcctacaggc
     1801 gatggcacgc taagtgctat gtcgacagcc taagactaca aaagaaaatg aggttggagt
     1861 gggaaaaaga ggaagaactt aagaagatcc gagagaagga agaatggata cgaatggact
     1921 attaccggag gcacaatcct aaaacaaccg aagactttga gctccttttc aatgctctag
     1981 aactttggcg tcaagaagaa gtcacacaag ttgagcaata ttcaagtgag gctgaaagga
     2041 aagctgctct gtgcgagctt ttggaaaaag agacgcagat aattgcttcc atcgggagac
     2101 atagatacgt tgctcagttg gagagacagg atgtagctac acaagctttt ctggataagt
     2161 gttcagcttc aaatacctgg agaagacctg atggacaaac cgttgaaatg gacacacagt
     2221 tcacaataag agccagagag ctgcaaagca tctacaagtg tatcctgctg aaaaacctct
     2281 ctcaagatga gaggctggat atactcctga cactgaagca cactgtgaag gaacatgagt
     2341 gtaaactgac ccaagaaatc ctagaattga ttgacagaga agttgacctt atgatgagag
     2401 gagttaaatc tcataatctc gaaggactca ggaaaagaat cacaacactc tttattcact
     2461 atatcaaaac tcctctgttt aacccagaag tcgcaaaaca ccttaaggtc cctcaagacc
     2521 cactgaagtt ttatgataag atttacttct gccacagttg ccaaatgtat ttgccttctg
     2581 tcgagttttc cgtctcaccc acctcgcatc gtgtgtaccg ctgccggcac tgcataaacc
     2641 tggagaatga gactcagagg cgggaatcat ttttgaaata caagtgttta cttcagcggc
     2701 tctacttctc agaagccaat tatgaggata actcccacat tgcttatcta atgcagcttc
     2761 aggatattaa gtacttgaca gagaacatct gggcatcgca gtcggccctc agcgcctggg
     2821 atgacctcaa tgacctggtc atggtcaggt ggaacaaaca cgtggagtgg tcgccctgga
     2881 actgcattct gctcaccaag gatgaaggca cagctcatct caagctagag agcatcgaga
     2941 agggatatgg acgccttttt attcacaaga tcaagcacaa acatatcttg gctaaaaact
     3001 atttttcaca ggtcccagtg ctggcttcac ttatacttaa ggatgatgaa atagaagcta
     3061 tcagaagtaa atactccaca gagaatcctc ctaaaatcat agtaccccat gggattgtac
     3121 ctaatggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3181 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3241 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3301 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3361 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3421 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3481 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3541 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3601 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3661 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3721 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3781 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3841 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3901 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3961 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4021 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4081 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4201 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4261 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4321 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4441 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4501 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4561 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4621 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4741 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4801 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4861 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4921 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4981 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5041 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5101 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5161 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5221 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5281 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5341 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5401 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5461 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5521 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5581 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5641 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5701 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5761 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5821 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5881 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5941 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6001 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6061 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8281 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8341 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8401 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
