LOCUS       V045572                 7344 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045572
VERSION     V045572
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..2024
                     /label="Serpinb12(NM_027971)"
                     /note="Serpinb12(NM_027971)"
                     /gene="Serpinb12"
     CDS             2031..2096
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2120..2708
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2774..2822
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3024..3452
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3466..3795
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3862..4656
                     /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    4832..4953
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5002..5018)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5026..5042
                     /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(5050..5080)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5095..5116
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5404..5992)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6163..7023)
                     /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(7024..7128)
                     /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 ccaccatgga ctctcttact gcagcaaata
      781 ataaattttg cttcgatttt ttccgagaga ttagcaaaga tgatgctcat aaaaatatct
      841 tcgtctgccc tctgagtctc tcagctgcct ttggcatggt ccgcctggga gctcgagggg
      901 acagtgcaca tcagattgat gaggcattac atttcaatga actttctaag gatgaacaca
      961 aagaacccaa tgatccctct ccacaaagtg aaagcaaagc ttctgacagc tctctggaag
     1021 ggcagaaaca aacatcagct tctcaggatc agcagggtga atctacaaat gaccatcaat
     1081 tgcttggctg ccactttggg aaacttctct ccagaataga cagagacaaa tcttattaca
     1141 ctttaagtat ggccaacaga ctctatggag agcaggaatt tccaatctgc tcagaatact
     1201 ctgatgacgt aacggaattt ttccacacaa cagtcgaaag tgtagatttc cagaaggatt
     1261 ctgagaaatc aagacaagag attaacttct gggttgaaag tcaaagccaa ggtaaaatca
     1321 aggaactgtt tggcaaggag gctattgata actcaactgt gctggtgctg gtgaatgctg
     1381 tttacttcaa ggccaagtgg gagagagaat tcaactctga aaacaccgtt gatgcatctt
     1441 tctgtctcaa tgagaacgag aagaagactg taaagatgat gaatcagaag ggtaagttta
     1501 gaattggctt tatcgacgag ctgcaggcac agatcctgga aatgaagtat gccatgggga
     1561 agctcagcat gctggtcctg cttccatctt gctcagaaga caatgtgaat agcctgcaag
     1621 agcttgaaaa gaaaataaat catgaaaaac tactggcctg gagtagctca gaaaatttgt
     1681 cagaaaaacc tgtagctatt tcctttcccc agttcaacct ggaagatagc tatgatctca
     1741 agtcgattct acaggacatg ggcatcaagg atgtctttga tgaaacgaag gctgacctta
     1801 ctggaatctc taagagtccc aatctgtatt tgtccaaaat tgtccacaag acctttgtgg
     1861 aggtggatga gatgggtacc caggcagctg cagccagtgg ggttgtggct gcagagaagg
     1921 cccttccatc ttgggtggag tttaatgcca accacccttt cctctttttt atccgtcaca
     1981 atccaaccca gagtttactt ttctgtggca gagtctattg ccctggatcc gactacaaag
     2041 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     2101 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2161 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2221 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2281 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2341 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2401 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2461 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2521 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2581 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2641 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2701 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     2761 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     2821 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     2881 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     2941 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3001 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3061 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3181 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3361 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     3421 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3541 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3601 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3721 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3781 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     4681 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4741 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4801 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4861 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4921 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4981 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5041 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5101 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5161 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5221 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5281 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5341 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5401 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5461 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5521 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5581 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5641 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5701 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5761 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5821 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5881 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5941 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6001 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6061 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6121 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6241 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6301 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6361 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6421 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6481 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6541 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6601 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6661 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6721 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6781 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6841 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6901 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6961 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7021 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7081 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7141 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     7201 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7261 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7321 aattgcatga agaatctgct tagg
//
