LOCUS       V023307                 8244 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023307
VERSION     V023307
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..2924
                     /label="Trim42(NM_001013955)"
                     /note="Trim42(NM_001013955)"
                     /gene="Trim42"
     CDS             2931..2996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3020..3608
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3674..3722
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3924..4352
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4366..4695
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4762..5556
                     /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    5732..5853
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5902..5918)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5926..5942
                     /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(5950..5980)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5995..6016
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6304..6892)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7063..7923)
                     /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(7924..8028)
                     /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 ctcggatccg ccaccatgga cacagctatg tgtgtctgtt
      781 ctccatgctg cacctggcag agatgctgcc cccggctgtt ctcctgttta tgctgcaagt
      841 tcatcttcac ctcggagcgg aactgcacct gcttcccctg cccctacaag gatgaacgga
      901 actgccagtt ctgccactgc acctgcgcgg agaaccccaa ttgccattgg tgctgctgct
      961 cctgggccaa tgaccccaac tgtaagtgct gctgcacagc cagcaccaat ctgaagtgct
     1021 actactatga gagccgctgc tgccgcaatg tcaccgtcac atttagaaag ggtcgccttc
     1081 gaagcatccg cacctcctca aagaccgccc tccgcgtggg gagcagcgac actcagcttg
     1141 atgagcccaa gaccatgcct gccagcagcc acctggtgaa ccacctcacc tgccccatgt
     1201 gcaaccggct gcgcctgcac tccttcatgc tgccctgcaa ccacagcctg tgcgagaagt
     1261 gcctgaggca actgcaaaaa cacgctgagg tcactgagaa tttcttcatc ctcatctgcc
     1321 ccatgtgtaa ccgctcgcac tgcatgccct acagccacca gatgcacctg ccagaaaact
     1381 acctgcgcgg gcgcctcacc aaacgctaca tgcagcagca tggctacctc aagtggcgct
     1441 tcgaccggtc ctcgggaccc atcctctgcc aagtgtgccg cacccggcgc attgcctaca
     1501 agcgttgtgt tacctgtcgc ctcaacctgt gcaatgactg tctcaaggcc ttccactcgg
     1561 atgtggccat gcaggatcac gtcttcgtgg acaccagcgc cgaggatcag gatgagaaga
     1621 tttgcatcca ccatccatcc agccgcatca acgagtactg tcgcagcgac aaccagctac
     1681 tctgcgcctt ctgcaagatc gctttccaca atggccatga cacggtcagc ctcatagacg
     1741 cctgctccga gaggtctgca gccctcttca gcgccatcgc caagttcaaa gcagtccgat
     1801 atgagattga caatgacctg atggaattca acattctgaa gagtagcttc aaagctgaca
     1861 aggaagcaaa gcggaaggag gtccggaatg gattcctcaa actgcgagcc atccttcaag
     1921 agaaagagaa gatcatcatg gaacagatcg agaacctaga agtgtccagg cagaaagaaa
     1981 ttgagaaata tgtatacatc acaaccatga aagtgaacga aatggatggt ctgattgcct
     2041 attccaagga agccctgaag gaaacaggcc aggtggcgtt cctgcagtct gccaagatcc
     2101 tggtggacca gattgaggag ggcatccaga ataccttcag gcctgaccct cagctgcggc
     2161 ttcactccct acactgtatt cccttggact ttgcagaact ctccagtgcc atccacgagc
     2221 tttttcccac aggacccaaa aagacttgct cttcagggga ctcactccct tcccagtatc
     2281 ctgtccactc ggaaatgatg atcgccagga aggtcacatt cagcacccac agcttcggca
     2341 accagcagat ataccagcgg agctcttcct tgatatcctt caacaccgcc aacgacaagg
     2401 gcaaggcggg tctagagaac tatgggagag cccagtctgc cgcacctgcc aaaacaactg
     2461 atggccttta tacttactgg agtgccacag gggaaaccca gcctccgcag agtagcaaca
     2521 gcttccacaa ctggtattct ttcaacgaca cctctgtgag gaccccaggc ccaattgtca
     2581 tctaccagac tctggtatac ccaagagcgg ccaaggttta ctggacgtgc ccaacagaag
     2641 atgtggactc ttttgagatg gaattctatg agctcatcac cacccctcct aataatgttc
     2701 gcacagagct ctgtggacaa attcgagaca tcatgcagca gaacctggag ctgcacaacc
     2761 tgacccccaa caccgagtat ttgtttaagg ttagagccat caatgaaaat ggtcccgggc
     2821 agtggagtga catctgcaag gtggtaacac cagatgggcg tgggaagaac cgagctaagt
     2881 ggggactgct gaagaatatc cagtctgccc tccagaagcg cttcctcgag gactacaaag
     2941 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     3001 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3061 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3121 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3181 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3241 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3301 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3361 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3421 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3481 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3541 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3601 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3661 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     3721 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3781 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3841 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3901 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4081 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4141 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4201 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4261 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4321 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4441 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4501 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4561 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4621 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4681 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4741 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4801 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4861 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4921 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4981 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5041 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5101 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5161 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5221 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5281 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5341 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5401 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5461 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5521 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5581 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5641 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5701 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5761 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5821 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5881 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5941 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6001 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6061 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6121 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6181 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6481 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6721 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7201 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8041 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8101 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8161 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8221 aattgcatga agaatctgct tagg
//
