LOCUS       V035876                 8935 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035876
VERSION     V035876
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..3594
                     /label="IREB2(NM_004136)"
                     /note="IREB2(NM_004136)"
                     /gene="IREB2"
     CDS             3601..3627
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3631..3657
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3661..3687
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3711..4299
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4365..4413
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4615..5043
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5057..5386
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5453..6247
                     /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    6423..6544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6593..6609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6617..6633
                     /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(6641..6671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6686..6707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6995..7583)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7754..8614)
                     /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(8615..8719)
                     /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 ccaccatgga cgccccaaaa
      721 gcaggatacg cctttgagta ccttattgaa acattaaatg acagttcaca taagaagttc
      781 ttcgatgtat ctaaacttgg caccaagtat gatgttctgc cttactcaat acgggtcttg
      841 ttggaagctg ctgtacgaaa ttgtgatggc tttttaatga agaaggaaga tgttatgaac
      901 attttagact ggaaaaccaa acaaagcaat gttgaagtgc cctttttccc tgcccgtgtt
      961 cttcttcaag attttactgg aataccagca atggtggatt ttgctgctat gagggaggca
     1021 gtgaaaactc ttggaggtga tcctgagaaa gtccatcctg cttgtccgac agatcttaca
     1081 gttgaccatt ctttacaaat tgacttcagt aaatgtgcaa tacagaatgc accaaatcct
     1141 ggaggtggtg acctgcagaa agcaggaaag ctctctccac ttaaagtgca gcctaagaag
     1201 cttccctgca gaggccagac tacctgccga ggatcttgtg attctggaga actaggccga
     1261 aactcaggaa cattttcttc gcagattgag aatacaccca tcctgtgtcc ttttcatttg
     1321 caaccagtgc ctgaacctga aacagtgtta aaaaatcaag aagtagaatt cggcagaaat
     1381 cgagagaggc ttcagttttt taagtggagt tcaagagttt ttaagaatgt ggcagtgatc
     1441 cctcctggaa ctggaatggc tcatcaaata aacttagaat atttgtcaag agtggttttt
     1501 gaagaaaaag acctcctctt cccagacagt gtagtcggca cagattcaca cataacgatg
     1561 gtgaatggtt tagggattct ggggtggggg gttggaggca ttgaaacaga agcagttatg
     1621 cttggtctgc cagtttctct tactttacca gaggtggttg gatgtgagtt aactgggtca
     1681 tcaaaccctt ttgttacatc catagatgtt gttcttggta ttacaaagca cctcaggcaa
     1741 gtaggagtgg ctggaaagtt tgttgagttt tttggaagtg gagtttcaca attatctata
     1801 gttgatcgaa ctacaatagc aaacatgtgt ccggaatatg gtgctatcct cagctttttc
     1861 cctgttgaca atgtgacatt aaaacattta gaacatacag gttttagcaa agccaaactc
     1921 gaatcaatgg aaacatacct taaagctgtg aaattgtttc gaaatgacca gaattcttca
     1981 ggagaacctg aatactccca ggtgatccag attaatctga attcaatagt tccatctgtt
     2041 agtggtccaa aaagacctca ggatagagtt gctgtgacag atatgaaaag cgatttccag
     2101 gcttgcttaa atgaaaaggt tggatttaaa ggcttccaaa ttgcagctga aaaacaaaag
     2161 gatattgtct ccattcatta tgaaggaagt gaatataagc tgtctcatgg atcagtggtc
     2221 attgctgcag ttatcagttg taccaataat tgcaatccat ctgtcatgct tgctgcaggt
     2281 cttttggcta aaaaggctgt tgaagctggt ctgcgtgtta aaccttatat aagaacaagt
     2341 ttatctccag gcagtgggat ggttacacat tacctcagtt caagtggagt attaccatat
     2401 ctaagtaagc ttggatttga aatcgttggc tatggatgtt caatttgtgt gggaaataca
     2461 gcacccttat cagacgcagt tttaaatgca gtaaaacagg gtgatttggt tacctgtgga
     2521 attttatctg gaaacaaaaa ttttgaaggt cgtctttgtg attgtgttcg tgccaattat
     2581 cttgcctctc cacccttagt ggtagcttat gccatagcag gcacagtgaa tatagatttc
     2641 cagacagaac ctttaggtac tgaccccacc ggcaagaaca tttacctgca tgatatttgg
     2701 cctagtcgag aagaagttca tcgagtagag gaagaacatg ttatactatc catgtttaaa
     2761 gcattaaaag ataaaataga aatggggaat aaacggtgga attccttaga agcaccggat
     2821 tcagttttgt ttccatggga cttaaagtct acttatatca gatgcccttc attttttgat
     2881 aaacttacca aagagccaat tgcactccag gctattgaaa atgcccatgt cttattatat
     2941 ttgggagact ctgtcacaac agatcatata tcacctgcag gaagtatcgc taggaatagt
     3001 gctgccgcta agtatttgac aaacagaggc cttacccctc gtgaattcaa ctcttacgga
     3061 gctcgaagag gtaatgatgc tgtaatgaca agaggcactt ttgcaaatat caagcttttt
     3121 aataagttta ttggaaaacc agctcctaaa acaattcatt ttccatcagg acagacgcta
     3181 gatgtatttg aggctgcaga gctgtaccag aaagaaggta tcccactgat tattttagca
     3241 ggaaagaaat atggttcagg aaactccaga gactgggctg ccaaaggacc gtatttactg
     3301 ggtgtgaaag ctgttttggc cgaaagttat gaaaaaatac acaaagatca tttgattgga
     3361 attggcatag ctccacttca gttccttcca ggagaaaatg cagattcctt gggcctctcc
     3421 ggtagagaaa cattttcttt aacatttcct gaagaactgt ctcctggaat tacattgaat
     3481 atacagacaa gcactggaaa agtattcagc gtgattgctt cgtttgaaga tgatgtggaa
     3541 ataacattat acaaacatgg aggattatta aactttgtgg cacgaaaatt ctcactcgag
     3601 tacccatacg acgtacctga ttatgcaact tatccttatg atgtacctga ttatgctagc
     3661 tatccttatg atgtcccaga ctacgcatag tgagggcccg atatctcgac aatcaacctc
     3721 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3781 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3841 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3901 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3961 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4021 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4081 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4141 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4201 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4261 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac
     4321 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4381 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4441 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4501 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4561 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     4621 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4681 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4741 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4801 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4861 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4921 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4981 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5041 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5161 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5221 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5281 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5341 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5401 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5461 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5521 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5581 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5641 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5701 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5761 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5821 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5881 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5941 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6001 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6061 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6121 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6181 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6241 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     6301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7501 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8761 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     8821 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     8881 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
