LOCUS       V031941                 7936 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031941
VERSION     V031941
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"
     CDS             706..2595
                     /label="TRIM25(NM_005082)"
                     /note="TRIM25(NM_005082)"
                     /gene="TRIM25"
     CDS             2602..2628
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2632..2658
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2662..2688
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2712..3300
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3366..3414
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3616..4044
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4058..4387
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4454..5248
                     /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    5424..5545
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5594..5610)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5618..5634
                     /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(5642..5672)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5687..5708
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5996..6584)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6755..7615)
                     /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(7616..7720)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc agagctgtgc
      721 cccctggccg aggagctgtc gtgctccatc tgcctggagc ccttcaagga gccggtcacc
      781 actccgtgcg gccacaactt ctgcgggtcg tgcctgaatg agacgtgggc agtccagggc
      841 tcgccatacc tgtgcccgca gtgccgcgcc gtctaccagg cgcgaccgca gctgcacaag
      901 aacacggtgc tgtgcaacgt ggtggagcag ttcctgcagg ccgacctggc ccgggagcca
      961 cccgccgacg tctggacgcc gcccgcccgc gcctctgcac ccagcccgaa tgcccaggtg
     1021 gcctgcgacc actgcctgaa ggaggccgcc gtgaagacgt gcttggtgtg catggcctcc
     1081 ttctgtcagg agcacctgca gccgcacttc gacagccccg ccttccagga ccacccgctg
     1141 cagccgcccg ttcgcgacct gttgcgccgc aaatgttccc agcacaatcg gctgcgggaa
     1201 tttttctgcc ccgagcacag cgagtgcatc tgccacatct gcctggtgga gcataagacc
     1261 tgctctcccg cgtccctgag ccaggccagc gccgacctgg aggccaccct gaggcacaaa
     1321 ctaactgtca tgtacagtca gatcaacggg gcgtcgagag cactggatga tgtgagaaac
     1381 aggcagcagg atgtgcggat gactgcaaac agaaaggtgg agcagctaca acaagaatac
     1441 acggaaatga aggctctctt ggacgcctca gagaccacct cgacaaggaa gataaaggaa
     1501 gaggagaaga gggtcaacag caagtttgac accatttatc agattctcct caagaagaag
     1561 agtgagatcc agaccttgaa ggaggagatt gaacagagcc tgaccaagag ggatgagttc
     1621 gagtttctgg agaaagcatc aaaactgcga ggaatctcaa caaagccagt ctacatcccc
     1681 gaggtggaac tgaaccacaa gctgataaaa ggcatccacc agagcaccat agacctcaaa
     1741 aacgagctga agcagtgcat cgggcggctc caggagccca cccccagttc aggtgaccct
     1801 ggagagcatg acccagcgtc cacacacaaa tccacacgcc ctgtgaagaa ggtctccaaa
     1861 gaggaaaaga aatccaagaa acctccccct gtccctgcct tacccagcaa gcttcccacg
     1921 tttggagccc cggaacagtt agtggattta aaacaagctg gcttggaggc tgcagccaaa
     1981 gccaccagct cacatccgaa ctcaacatct ctcaaggcca aggtgctgga gaccttcctg
     2041 gccaagtcca gacctgagct cctggagtat tacattaaag tcatcctgga ctacaacacc
     2101 gcccacaaca aagtggctct gtcagagtgc tatacagtag cttctgtggc tgagatgcct
     2161 cagaactacc ggccgcatcc ccagaggttc acatactgct ctcaggtgct gggcctgcac
     2221 tgctacaaga aggggatcca ctactgggag gtggagctgc agaagaacaa cttctgtggg
     2281 gtaggcatct gctacggaag catgaaccgg cagggcccag aaagcaggct cggccgcaac
     2341 agcgcctcct ggtgcgtgga gtggttcaac accaagatct ctgcctggca caataacgtg
     2401 gagaaaaccc tgccctccac caaggccacg cgggtgggcg tgcttctcaa ctgtgaccac
     2461 ggctttgtca tcttcttcgc tgttgccgac aaggtccacc tgatgtataa gttcagggtg
     2521 gactttactg aggctttgta cccggctttc tgggtatttt ctgctggtgc cacactctcc
     2581 atctgctccc ccaagctcga gtacccatac gacgtacctg attatgcaac ttatccttat
     2641 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata gtgagggccc
     2701 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2761 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2821 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2881 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2941 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3001 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3061 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     3121 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3181 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3241 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3301 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     3361 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     3421 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     3481 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     3541 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     3601 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3661 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3721 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     3781 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3841 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3901 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3961 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     4021 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4081 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4141 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4261 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4321 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4381 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4441 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4501 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4561 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4621 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4681 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4741 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4801 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4861 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4921 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4981 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5041 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5101 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5161 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5221 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     5281 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5341 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5401 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5461 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5521 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5581 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5641 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5701 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5761 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5821 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5881 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5941 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6001 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6061 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6121 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6181 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6241 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6301 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6361 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6421 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6481 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6541 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6601 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6661 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6721 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6781 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6841 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6901 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6961 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7021 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7081 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7141 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7201 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7261 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7321 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7381 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7441 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7501 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7561 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7621 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7681 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7741 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     7801 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7861 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7921 gaagaatctg cttagg
//
