LOCUS       V023637                 7394 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023637
VERSION     V023637
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             779..2074
                     /label="PAG1(NM_018440)"
                     /note="PAG1(NM_018440)"
                     /gene="PAG1"
     CDS             2081..2146
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2170..2758
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2824..2872
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3074..3502
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3516..3845
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3912..4706
                     /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    4882..5003
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5052..5068)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5076..5092
                     /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(5100..5130)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5145..5166
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5454..6042)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6213..7073)
                     /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(7074..7178)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggggcccgcg gggagcctgc tgggcagcgg acagatgcag atcaccctgt ggggaagtct
      841 ggctgctgtc gccattttct tcgtcatcac cttcctcatc ttcctgtgct ctagttgtga
      901 cagggaaaag aagccgcgac agcatagtgg ggaccatgag aacctgatga acgtgccttc
      961 agacaaggag atgttcagcc gttcagttac tagcctggca acagatgctc ctgccagcag
     1021 tgagcagaat ggggcactca ccaatgggga cattctttca gaggacagta ctctgacctg
     1081 catgcagcat tacgaggaag tccagacatc ggcctcggat ctgctggatt cccaggacag
     1141 cacagggaaa ccaaaatgtc atcagagtcg ggagctgccc agaatccctc ccgagagcgc
     1201 agtggatacc atgctcacgg cgagaagtgt ggacggggac caggggctgg ggatggaagg
     1261 gccctatgaa gtgctcaagg acagctcctc ccaagaaaac atggtggagg actgcttgta
     1321 tgaaactgtg aaagagatca aggaggtggc tgcagctgca cacctggaga aaggccacag
     1381 tggcaaggca aaatctactt ctgcctcgaa agagctccca gggccccaga ctgaaggcaa
     1441 agctgagttt gctgaatatg cctcggtgga cagaaacaaa aaatgtcgtc aaagtgttaa
     1501 tgtagagagt atccttggaa attcatgtga tccagaagag gaggccccac cacctgtccc
     1561 tgttaagctt ctggacgaga atgaaaacct tcaggagaag gaagggggag aggcggaaga
     1621 gagtgccaca gacacgacca gtgaaactaa caagagattt agctcattgt catacaagtc
     1681 tcgggaagaa gaccccactc tcacagaaga agagatctca gctatgtact catcagtaaa
     1741 taaacctgga cagttagtga ataaatcggg gcagtcgctt acagttccgg agtccaccta
     1801 cacctccatt caaggggacc cacagaggtc accctcctcc tgtaatgatc tctatgctac
     1861 tgttaaagac ttcgaaaaaa ctccaaacag cacacttcca ccagcaggga ggcccagcga
     1921 ggagccagag cctgattatg aagcgataca gactctcaac agagaggaag aaaaggccac
     1981 cctggggacc aatggccacc acggtctcgt cccaaaggag aacgactacg agagcataag
     2041 tgacttgcag caaggcagag atattaccag gctcctcgag gactacaaag accatgacgg
     2101 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2161 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2221 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2281 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2341 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2401 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2461 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2521 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2581 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2641 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2701 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2761 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     2821 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2881 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     2941 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3001 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3061 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3421 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3481 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3781 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3841 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3901 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3961 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4021 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4081 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4141 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4201 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4261 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4321 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4381 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4441 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4501 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4561 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4621 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4681 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4741 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4801 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4861 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4981 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5041 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5641 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5881 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7201 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7261 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7321 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7381 agaatctgct tagg
//
