LOCUS       V045439                 7314 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045439
VERSION     V045439
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..1994
                     /label="Iigp1(NM_021792)"
                     /note="Iigp1(NM_021792)"
                     /gene="Iigp1"
     CDS             2001..2066
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2090..2678
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2744..2792
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2994..3422
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3436..3765
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3832..4626
                     /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    4802..4923
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4972..4988)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4996..5012
                     /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(5020..5050)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5065..5086
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5374..5962)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6133..6993)
                     /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(6994..7098)
                     /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 ccaccatggg tcagctgttc tcttcaccta
      781 agagtgatga gaataatgat ttgccctcca gctttactgg ttattttaag aaatttaata
      841 cgggaagaaa aatcatttct caagagatcc tcaatttgat tgaattaagg atgagaaaag
      901 ggaatattca gttgacaaac tctgcaatca gtgatgcatt aaaagaaatc gatagtagtg
      961 tgctcaatgt tgctgtcacc ggggagacgg gatcagggaa gtccagcttc atcaataccc
     1021 tgagaggcat tgggaatgaa gaagaaggtg cagctaaaac tggggtggtg gaggtaacca
     1081 tggaaagaca tccatacaaa caccccaata tacccaatgt ggttttttgg gacctgcctg
     1141 ggattggaag cacaaatttc ccaccaaaca cttacctgga gaaaatgaag ttctatgagt
     1201 acgatttctt cattattatt tcggccacac gcttcaagaa aaatgatata gacattgcca
     1261 aagcaatcag catgatgaag aaggaattct acttcgtgag aaccaaggtg gactctgaca
     1321 taacaaatga agcagatggc aaacctcaaa cctttgacaa agaaaaggtc ctgcaggaca
     1381 tccgccttaa ctgtgtgaac acctttaggg agaatggcat tgctgagcca ccaatcttcc
     1441 tgctctctaa caaaaatgtt tgtcactatg acttccccgt cctgatggac aagctgataa
     1501 gtgacctccc tatctacaag agacacaatt ttatggtctc cttacccaat atcacagatt
     1561 cagtcattga aaagaagcgg caatttctga agcagaggat ttggctggaa ggatttgctg
     1621 ctgacctagt gaatatcatc ccttctctga cctttctctt ggacagtgat ttggagactc
     1681 tgaagaaaag catgaaattc taccgcactg tgtttggagt ggatgaaaca tctttgcaga
     1741 gattagctag ggactgggaa atagaggtgg atcaggtgga ggccatgata aaatctcctg
     1801 ctgtgttcaa acctacagat gaagaaacaa tacaagaaag gctttcaaga tatattcagg
     1861 agttctgttt ggctaatggg tacttacttc ctaaaaatag ttttcttaaa gaaatatttt
     1921 acctgaaata ttatttcctt gacatggtga ctgaggatgc taaaactctt cttaaagaga
     1981 tatgtttaag aaacggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     2041 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     2101 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2161 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2221 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2281 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2341 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2401 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2461 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     2521 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2581 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2641 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     2701 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     2761 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     2821 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     2881 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     2941 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3001 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3061 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3121 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3361 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3421 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3481 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3541 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3601 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3661 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3721 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3781 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3841 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3901 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3961 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4021 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4081 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4141 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4201 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4261 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4321 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4381 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4441 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4501 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4561 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4621 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     4681 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4741 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4801 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4861 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4921 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4981 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5041 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5101 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5161 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5221 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5281 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5341 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5401 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5461 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5521 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     5581 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5641 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5701 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5761 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5821 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5881 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5941 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6001 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6061 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7081 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7141 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7201 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7261 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
