LOCUS       V041859                 6038 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041859
VERSION     V041859
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"
     CDS             941..1570
                     /label="Cldn4(NM_009903)"
                     /note="Cldn4(NM_009903)"
                     /gene="Cldn4"
     CDS             1577..1600
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    1638..1862
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      1908..2336
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2350..2679
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2746..3540
                     /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    3714..3835
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3884..3900)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3908..3924
                     /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(3932..3962)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3977..3998
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4286..4871)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5042..5902)
                     /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(5903..6007)
                     /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 aagcttggta ccgagctcgg atccgccacc atggcgtcta tgggactaca
      961 ggtcctggga atctccttgg cagtcctggg ctggctgggg atcatcctga gttgtgcgct
     1021 ccccatgtgg cgggtgaccg ccttcatcgg cagcaacatc gtcacggcac agaccagctg
     1081 ggagggcctc tggatgaact gcgtggtgca gagcacaggt cagatgcagt gcaagatgta
     1141 cgactcgatg ctcgccctgc cgcaggacct gcaggccgcc cgagccctta tggtcatcag
     1201 catcatcgtg ggtgctctgg ggatgcttct ctcagtggta gggggcaagt gcaccaactg
     1261 catggaggac gagaccgtca aggccaagat catgatcacc gccggagccg tgttcatcgt
     1321 ggcaagcatg ctgattatgg tgcccgtgtc ctggaccgct cacaacgtca tccgcgactt
     1381 ctacaaccct atggtggctt ccgggcagaa gagggaaatg ggggcctcgc tttacgtcgg
     1441 ctgggcggcc tccgggctgc tgctcctggg aggaggcctc ctctgctgca gttgcccacc
     1501 tcgtagcaac gacaagccct actcggccaa gtactccgcc gcccgctctg tccccgccag
     1561 caactatgtg ctcgaggatt acaaggatga cgacgataag tagtgagggc ccgtttaaac
     1621 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1681 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1741 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1801 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1861 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1921 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1981 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2041 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2101 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2161 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2221 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2281 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2341 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2401 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2461 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2521 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2581 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2641 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2701 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2761 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2821 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2881 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2941 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3001 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3061 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3121 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3181 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3241 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3301 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3361 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3421 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3481 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3541 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3601 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3661 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3721 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3781 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3841 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3901 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3961 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4021 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4081 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4141 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4201 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4261 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4321 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4381 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4441 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4501 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4561 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4621 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4681 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4741 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4801 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4861 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4921 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4981 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5041 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5101 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5161 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5221 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5281 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5341 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5401 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5461 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5521 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5581 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5641 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5701 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5761 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5821 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5881 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5941 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6001 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
