LOCUS       V039236                 8755 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039236
VERSION     V039236
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..3414
                     /label="ITCH(NM_001257137)"
                     /note="ITCH(NM_001257137)"
                     /gene="ITCH"
     CDS             3421..3447
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3451..3477
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3481..3507
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3531..4119
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4185..4233
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4435..4863
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4877..5206
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5273..6067
                     /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    6243..6364
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6413..6429)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6437..6453
                     /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(6461..6491)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6506..6527
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6815..7403)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7574..8434)
                     /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(8435..8539)
                     /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 ccaccatgtc tgacagtgga
      721 tcacaacttg gttcaatggg tagcctcacc atgaaatcac agcttcagat cactgtcatc
      781 tcagcaaaac ttaaggaaaa taagaagaat tggtttggac caagtcctta cgtagaggtc
      841 acagtagatg gacagtcaaa gaagacagaa aaatgcaaca acacaaacag tcccaagtgg
      901 aagcaacccc ttacagttat cgttacccct gtgagtaaat tacattttcg tgtgtggagt
      961 caccagacac tgaaatctga tgttttgttg ggaactgctg cattagatat ttatgaaaca
     1021 ttaaagtcaa acaatatgaa acttgaagaa gtagttgtga ctttgcagct tggaggtgac
     1081 aaagagccaa cagagacaat aggagacttg tcaatttgtc ttgatgggct acagttagag
     1141 tctgaagttg ttaccaatgg tgaaactaca tgttcagaaa atggggtttc gctctgtttg
     1201 cccaggctgg agtgcaatag tgcgatctcg gctcactgca acctctgcct cccgggttta
     1261 agtgattctc ctatctcagc ctcccgagta gctgggttta caggtgcttc tcagaatgat
     1321 gatggctcca gatccaagga tgaaacaaga gtgagcacaa atggatcaga tgaccctgaa
     1381 gatgcaggag ctggtgaaaa taggagagtc agtgggaata attctccatc actctcaaat
     1441 ggtggtttta aaccttctag acctccaaga ccttcacgac caccaccacc caccccacgt
     1501 agaccagcat ctgtcaatgg ttcaccatct gccacttctg aaagtgatgg gtctagtaca
     1561 ggctctctgc cgccgacaaa tacaaataca aatacatctg aaggagcaac atctggatta
     1621 ataattcctc ttactatatc tggaggctca ggccctaggc cattaaatcc tgtaactcaa
     1681 gctcccttgc cacctggttg ggagcagaga gtggaccagc acgggcgagt ttactatgta
     1741 gatcatgttg agaaaagaac aacatgggat agaccagaac ctctacctcc tggctgggaa
     1801 cggcgggttg acaacatggg acgtatttat tatgttgacc atttcacaag aacaacaacg
     1861 tggcagaggc caacactgga atccgtccgg aactatgaac aatggcagct acagcgtagt
     1921 cagcttcaag gagcaatgca gcagtttaac cagagattca tttatgggaa tcaagattta
     1981 tttgctacat cacaaagtaa agaatttgat cctcttggtc cattgccacc tggatgggag
     2041 aagagaacag acagcaatgg cagagtatat ttcgtcaacc acaacacacg aattacacaa
     2101 tgggaagacc ccagaagtca aggtcaatta aatgaaaagc ccttacctga aggttgggaa
     2161 atgagattca cagtggatgg aattccatat tttgtggacc acaatagaag aactaccacc
     2221 tatatagatc cccgcacagg aaaatctgcc ctagacaatg gacctcagat agcctatgtt
     2281 cgggacttca aagcaaaggt tcagtatttc cggttctggt gtcagcaact ggccatgcca
     2341 cagcacataa agattacagt gacaagaaaa acattgtttg aggattcctt tcaacagata
     2401 atgagcttca gtccccaaga tctgcgaaga cgtttgtggg tgatttttcc aggagaagaa
     2461 ggtttagatt atggaggtgt agcaagagaa tggttctttc ttttgtcaca tgaagtgttg
     2521 aacccaatgt attgcctgtt tgaatatgca gggaaggata actactgctt gcagataaac
     2581 cccgcttctt acatcaatcc agatcacctg aaatattttc gttttattgg cagatttatt
     2641 gccatggctc tgttccatgg gaaattcata gacacgggtt tttctttacc attctataag
     2701 cgtatcttga acaaaccagt tggactcaag gatttagaat ctattgatcc agaattttac
     2761 aattctctca tctgggttaa ggaaaacaat attgaggaat gtgatttgga aatgtacttc
     2821 tccgttgaca aagaaattct aggtgaaatt aagagtcatg atctgaaacc taatggtggc
     2881 aatattcttg taacagaaga aaataaagag gaatacatca gaatggtagc tgagtggagg
     2941 ttgtctcgag gtgttgaaga acagacacaa gctttctttg aaggctttaa tgaaattctt
     3001 ccccagcaat atttgcaata ctttgatgca aaggaattag aggtcctttt atgtggaatg
     3061 caagagattg atttgaatga ctggcaaaga catgccatct accgtcatta tgcaaggacc
     3121 agcaaacaaa tcatgtggtt ttggcagttt gttaaagaaa ttgataatga gaagagaatg
     3181 agacttctgc agtttgttac tggaacctgc cgattgccag taggaggatt tgctgatctc
     3241 atggggagca atggaccaca gaaattctgc attgaaaaag ttgggaaaga aaattggcta
     3301 cccagaagtc atacctgttt taatcgcctg gacctgccac catacaagag ctatgagcaa
     3361 ctgaaggaaa agctgttgtt tgccatagaa gaaacagaag gatttggaca agagctcgag
     3421 tacccatacg acgtacctga ttatgcaact tatccttatg atgtacctga ttatgctagc
     3481 tatccttatg atgtcccaga ctacgcatag tgagggcccg atatctcgac aatcaacctc
     3541 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3601 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3661 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3721 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3781 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3841 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3901 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     3961 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4021 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4081 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac
     4141 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4201 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4261 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4321 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4381 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     4441 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4501 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4561 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4621 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4681 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4741 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4801 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4861 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4921 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4981 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5041 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5101 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5161 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5221 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5281 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5341 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5401 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5461 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5521 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5581 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5641 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5701 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5761 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5821 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5881 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5941 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6001 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6061 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     6121 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6181 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6241 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6361 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6421 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6481 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6541 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8581 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     8641 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     8701 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
