LOCUS       V047352                 9082 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047352
VERSION     V047352
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1033..4602
                     /label="TJP2(NM_004817)"
                     /note="TJP2(NM_004817)"
                     /gene="TJP2"
     polyA_signal    4682..4906
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4952..5380
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5394..5723
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5790..6584
                     /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    6758..6879
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6928..6944)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6952..6968
                     /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(6976..7006)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7021..7042
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7330..7915)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8086..8946)
                     /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(8947..9051)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttatgccggt gcgaggagac cgcgggtttc caccccggcg ggagctgtca
     1081 ggttggctcc gcgccccagg catggaagag ctgatatggg aacagtacac tgtgacccta
     1141 caaaaggatt ccaaaagagg atttggaatt gcagtgtccg gaggcagaga caacccccac
     1201 tttgaaaatg gagaaacgtc aattgtcatt tctgatgtgc tcccgggtgg gcctgctgat
     1261 gggctgctcc aagaaaatga cagagtggtc atggtcaatg gcacccccat ggaggatgtg
     1321 cttcattcgt ttgcagttca gcagctcaga aaaagtggga aggtcgctgc tattgtggtc
     1381 aagaggcccc ggaaggtcca ggtggccgca cttcaggcca gccctcccct ggatcaggat
     1441 gaccgggctt ttgaggtgat ggacgagttt gatggcagaa gtttccggag tggctacagc
     1501 gagaggagcc ggctgaacag ccatgggggg cgcagccgca gctgggagga cagcccggaa
     1561 agggggcgtc cccatgagcg ggcccggagc cgggagcggg acctcagccg ggaccggagc
     1621 cgtggccgga gcctggagcg gggcctggac caagaccatg cgcgcacccg agaccgcagc
     1681 cgtggccgga gcctggagcg gggcctggac cacgactttg ggccatcccg ggaccgggac
     1741 cgtgaccgca gccgcggccg gagcattgac caggactacg agcgagccta tcaccgggcc
     1801 tacgacccag actacgagcg ggcctacagc ccggagtaca ggcgcggggc ccgccacgat
     1861 gcccgctctc ggggaccccg aagccgcagc cgcgagcacc cgcactcacg gagccccagc
     1921 cccgagccta gggggcggcc ggggcccatc ggggtcctcc tgatgaaaag cagagcgaac
     1981 gaagagtatg gtctccggct tgggagtcag atcttcgtaa aggaaatgac ccgaacgggt
     2041 ctggcaacta aagatggcaa ccttcacgaa ggagacataa ttctcaagat caatgggact
     2101 gtaactgaga acatgtcttt aacggatgct cgaaaattga tagaaaagtc aagaggaaaa
     2161 ctacagctag tggtgttgag agacagccag cagaccctca tcaacatccc gtcattaaat
     2221 gacagtgact cagaaataga agatatttca gaaatagagt caaaccgatc attttctcca
     2281 gaggagagac gtcatcagta ttctgattat gattatcatt cctcaagtga gaagctgaag
     2341 gaaaggccaa gttccagaga ggacacgccg agcagattgt ccaggatggg tgcgacaccc
     2401 actcccttta agtccacagg ggatattgca ggcacagttg tcccagagac caacaaggaa
     2461 cccagatacc aagaggaccc cccagctcct caaccaaaag cagccccgag aacttttctt
     2521 cgtcctagtc ctgaagatga agcaatatat ggccctaata ccaaaatggt aaggttcaag
     2581 aagggagaca gcgtgggcct ccggttggct ggtggcaatg atgtcgggat atttgttgct
     2641 ggcattcaag aagggacctc ggcggagcag gagggccttc aagaaggaga ccagattctg
     2701 aaggtgaaca cacaggattt cagaggatta gtgcgggagg atgccgttct ctacctgtta
     2761 gaaatcccta aaggtgaaat ggtgaccatt ttagctcaga gccgagccga tgtgtataga
     2821 gacatcctgg cttgtggcag aggggattcg ttttttataa gaagccactt tgaatgtgag
     2881 aaggaaactc cacagagcct ggccttcacc agaggggagg tcttccgagt ggtagacaca
     2941 ctgtatgacg gcaagctggg caactggctg gctgtgagga ttgggaacga gttggagaaa
     3001 ggcttaatcc ccaacaagag cagagctgaa caaatggcca gtgttcaaaa tgcccagaga
     3061 gacaacgctg gggaccgggc agatttctgg agaatgcgtg gccagaggtc tggggtgaag
     3121 aagaacctga ggaaaagtcg ggaagacctc acagctgttg tgtctgtcag caccaagttc
     3181 ccagcttatg agagggtttt gctgcgagaa gctggtttca agagacctgt ggtcttattc
     3241 ggccccatag ctgatatagc aatggaaaaa ttggctaatg agttacctga ctggtttcaa
     3301 actgctaaaa cggaaccaaa agatgcagga tctgagaaat ccactggagt ggtccggtta
     3361 aataccgtga ggcaaattat tgaacaggat aagcatgcac tactggatgt gactccgaaa
     3421 gctgtggacc tgttgaatta cacccagtgg ttcccaattg tgattttttt caacccagac
     3481 tccagacaag gtgtcaaaac catgagacaa aggttaaatc caacgtccaa caaaagttct
     3541 cgaaagttat ttgatcaagc caacaagctt aaaaaaacgt gtgcacacct ttttacagct
     3601 acaatcaacc taaattcagc caatgatagc tggtttggca gcttaaagga cactattcag
     3661 catcagcaag gagaagcggt ttgggtctct gaaggaaaga tggaagggat ggatgatgac
     3721 cccgaagacc gcatgtccta cttaaccgcc atgggcgcgg actatctgag ttgcgacagc
     3781 cgcctcatca gtgactttga agacacggac ggtgaaggag gcgcctacac tgacaatgag
     3841 ctggatgagc cagccgagga gccgctggtg tcgtccatca cccgctcctc ggagccggtg
     3901 cagcacgagg agagcataag gaaacccagc ccagagccac gagctcagat gaggagggct
     3961 gctagcagcg atcaacttag ggacaatagc ccgcccccag cattcaagcc agagccgccc
     4021 aaggccaaaa cccagaacaa agaagaatcc tatgacttct ccaaatccta tgaatataag
     4081 tcaaacccct ctgccgttgc tggtaatgaa actcctgggg catctaccaa aggttatcct
     4141 cctcctgttg cagcaaaacc tacctttggg cggtctatac tgaagccctc cactcccatc
     4201 cctcctcaag agggtgagga ggtgggagag agcagtgagg agcaagataa tgctcccaaa
     4261 tcagtcctgg gcaaagtcaa aatatttgag aagatggatc acaaggccag gttacagaga
     4321 atgcaggagc tccaggaagc acagaatgca aggatcgaaa ttgcccagaa gcatcctgat
     4381 atctatgcag ttccaatcaa aacgcacaag ccagaccctg gcacgcccca gcacacgagt
     4441 tccagacccc ctgagccaca gaaagctcct tccagacctt atcaggatac cagaggaagt
     4501 tatggcagtg atgccgagga ggaggagtac cgccagcagc tgtcagaaca ctccaagcgc
     4561 ggttactatg gccagtctgc ccgataccgg gacacagaat tataggaatt ctgcagatat
     4621 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     4681 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4741 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4801 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4861 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4921 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4981 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5041 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5101 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5161 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5221 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5281 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5341 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5641 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5701 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5761 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5821 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5881 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5941 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6001 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6061 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6121 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6181 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6241 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6301 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6361 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6421 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6481 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6541 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6601 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6661 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6721 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6781 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6841 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6901 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7741 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7861 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9061 ccgaaaagtg ccacctgacg tc
//
