LOCUS       V028581                 8005 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028581
VERSION     V028581
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..2664
                     /label="TRIM32(NM_012210)"
                     /note="TRIM32(NM_012210)"
                     /gene="TRIM32"
     CDS             2671..2697
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2701..2727
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2731..2757
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2781..3369
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3435..3483
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3685..4113
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4127..4456
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4523..5317
                     /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    5493..5614
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5663..5679)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5687..5703
                     /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(5711..5741)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5756..5777
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6065..6653)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6824..7684)
                     /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(7685..7789)
                     /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 tgcagcagca
      721 gcttctcacc tgaacctgga tgccctccgg gaagtgctag aatgccccat ctgcatggag
      781 tccttcacag aagagcagct gcgtcccaag cttctgcact gtggccatac catctgccgc
      841 cagtgcctgg agaagctatt ggccagtagc atcaatggtg tccgctgtcc cttttgcagc
      901 aagattaccc gcataaccag cttgacccag ctgacagaca atctgacagt gctaaagatc
      961 attgatacag ctgggctcag cgaggctgtg gggctgctca tgtgtcggtc ctgtgggcgg
     1021 cgtctgcccc ggcaattctg ccggagctgt ggtttggtgt tatgtgagcc ctgccgggag
     1081 gcagaccatc agcctcctgg ccactgtaca ctccctgtca aagaagcagc tgaggagcgg
     1141 cgtcgggact ttggagagaa gttaactcgt ctgcgggaac ttatggggga gctgcagcgg
     1201 cggaaggcag ccttggaagg tgtctccaag gaccttcagg caaggtataa agcagttctc
     1261 caggagtatg ggcatgagga gcgcagggtc caggatgagc tggctcgctc tcggaagttc
     1321 ttcacaggct ctttggctga agttgagaag tccaatagtc aagtggtaga ggagcagagt
     1381 tacctgctta acattgcaga ggtgcaggct gtgtctcgct gtgactactt cctggccaag
     1441 atcaagcagg cagatgtagc actactggag gagacagctg atgaggagga gccagagctc
     1501 actgccagct tgcctcggga gctcaccctg caagatgtgg agctccttaa ggtaggtcat
     1561 gttggccccc tccaaattgg acaagctgtt aagaagcccc ggacagttaa cgtggaagat
     1621 tcctgggcca tggaggccac agcgtctgct gcctctacct ctgttacttt tagagagatg
     1681 gacatgagcc cggaggaagt ggttgccagc cctagggcct cacctgctaa acagcggggt
     1741 cctgaggcag cctccaatat ccagcagtgc ctctttctca agaagatggg ggccaaaggc
     1801 agcactccag gaatgttcaa tcttccagtc agtctctacg tgaccagtca aggtgaagta
     1861 ctagtcgctg accgtggtaa ctatcgtata caagtcttta cccgcaaagg ctttttgaag
     1921 gaaatccgcc gcagccccag tggcattgat agctttgtgc taagcttcct tggggcagat
     1981 ctacccaacc tcactcctct ctcagtggca atgaactgcc aggggctgat tggtgtgact
     2041 gacagctatg ataactccct caaggtatat accttggatg gccactgcgt ggcctgtcac
     2101 aggagccagc tgagcaaacc atggggtatc acagccttgc catctggcca gtttgtagta
     2161 accgatgtgg aaggtggaaa gctttggtgt ttcacagttg atcgaggatc aggggtggtc
     2221 aaatacagct gcctatgtag tgctgtgcgg cccaaatttg tcacctgtga tgctgagggc
     2281 accgtctact tcacccaggg cttaggcctc aatctggaga atcggcagaa tgagcaccac
     2341 ctggagggtg gcttttccat tggctctgta ggccctgatg ggcagctggg tcgccagatt
     2401 agccacttct tctcggagaa tgaggatttc cgctgcattg ctggcatgtg tgtggatgct
     2461 cgtggtgatc tcatcgtggc tgacagtagt cgcaaggaaa ttctccattt tcctaagggt
     2521 gggggctata gtgtccttat tcgagaggga cttacctgtc cggtgggcat agccctaact
     2581 cctaaggggc agctgctggt cttggactgt tgggatcatt gcatcaagat ctacagctac
     2641 catctgagaa gatattccac cccactcgag tacccatacg acgtacctga ttatgcaact
     2701 tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga ctacgcatag
     2761 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2821 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2881 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2941 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3001 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3061 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3121 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3181 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3241 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3301 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3361 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3421 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3481 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3541 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3601 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3661 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3721 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3781 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3841 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3901 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3961 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4021 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     4081 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4141 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4201 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4321 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4381 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4441 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4501 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4561 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4621 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4681 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4741 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4801 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4861 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4921 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4981 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5041 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5101 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5161 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5221 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5281 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5341 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5401 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5461 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5521 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5581 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5641 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5701 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5761 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6241 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6481 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7801 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     7861 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7921 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7981 caattgcatg aagaatctgc ttagg
//
