LOCUS       V042771                 8427 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042771
VERSION     V042771
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        236..615
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        616..819
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        864..882
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             965..3106
                     /label="Cdh13(NM_019707)"
                     /note="Cdh13(NM_019707)"
                     /gene="Cdh13"
     CDS             3107..3172
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             3179..3232
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             3257..3976
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4028..4252
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4298..4726
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4740..5069
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5136..5930
                     /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    6104..6225
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6274..6290)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6298..6314
                     /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(6322..6352)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6367..6388
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6676..7261)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7432..8292)
                     /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(8293..8397)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc
      961 caccatgcag ccgagaactc cgctcaccct gtgcgtcctg ctgtcccagg tgctcctggt
     1021 cacgtctgca gatgatctgg agtgcacccc tggattccag cggaaagtgt tacacatcca
     1081 ccagcctgcc gaattcatcg aggaccagcc tgtcctaaac ttgaccttca atgactgcaa
     1141 gggcaacgag aagctgcact acgaggtctc aagcccacac ttcaaggtga acagcgacgg
     1201 caccttagtg gctctcagga acatcactgc ggtgggcagg accctgtttg tccatgcgag
     1261 gactcctcat gctgaagaca tggcagaact cgtgattgtc gggggcaaag acatccaggg
     1321 ctccttgcag gatatcttta aatttgcaag gacatctcct gtcccaagac aaaagaggtc
     1381 cattgtggtg tcccccatct tgatcccaga gaaccagaga cagcccttcc ccagagatgt
     1441 cggcaaggta gtcgatagcg acagacccga agggtccaag ttccggctca ccgggaaggg
     1501 agtggatcaa gaccctaaag gaactttcag aatcaatgag aacacaggca gtgtctccgt
     1561 gacacggacc ctggacagag aaacgatcgc tacttatcaa ctgtatgtgg aaaccacgga
     1621 tgccagtggc aaaactctgg aagggccagt gcctctggaa gtcattgtga ttgaccagaa
     1681 cgacaacaga cccatcttcc gggaaggccc ttacatcggc catgtcatgg aagggtcacc
     1741 cacagggacc acggtgatgc ggatgacagc gtttgatgca gatgacccgg ctactgacaa
     1801 tgctctcctg aggtacaaca tccgtcagca gacgcctgac aagccatctc ctaacatgtt
     1861 ctacatcgat cccgagaaag gagacattgt caccgtggtg tcacctgcgc tgctggaccg
     1921 ggagactctg gaaaacccca agtatgaact gatcatcgaa gctcaagata tggcaggatt
     1981 ggatgtcgga ctgacaggca cagccacagc caccatcgtg atcgatgaca aaaatgatca
     2041 ctcaccaaaa ttcaccaaga aagagtttca agccacagtg gaggagggag ctgtgggggt
     2101 cattgtcaac ttgacagtgg aagacaaaga cgaccctacc acaggagcct ggagggctgc
     2161 atacaccatc atcaatggaa atcctgggca gagcttcgag atccacacca acccacagac
     2221 caacgagggc atgctctctg tggtcaagcc cctggactat gagatctctg cctttcacac
     2281 cctgctgatc aaagtggaga atgaggaccc actggtaccc gacgtctcct atggccccag
     2341 ctccacggca actgtccaca tcacagtcct ggatgtcaac gagggaccag tcttctaccc
     2401 ggaccccatg atggtgacta aacaggagaa catctctgtg ggcagtgtgc tgctgacagt
     2461 gaatgccaca gacccagact ccctgcaaca tcaaactatc aggtactcta tctacaagga
     2521 cccagcaggc tggctgagta tcaaccccat caatggaacg gtggacacca cagctgtgtt
     2581 ggaccgggag tccccattcg tccacaacag tgtgtacacg gccctgttcc tggccatcga
     2641 cagtggcaac cctcctgcaa ctggcaccgg gactctgctg ataaccctag aggacattaa
     2701 tgacaacgct cctgtcattt acccaactgt ggctgaggtc tgcgatgatg ccagaaacct
     2761 cagcgtggtc attctggggg cttcggacaa ggaccttcac cccaacacag accccttcaa
     2821 gtttgagatt cataaacaga cagtccctga taaagtctgg aagatctcca aaatcaacaa
     2881 cacccacgcc ctcgtgagcc ttcttcagaa tctgaacaag gcaaactaca acctgcccat
     2941 catggtgaca gattcaggga agccacccat gacgaacatc acggacctca gggtgcaggt
     3001 gtgctcttgc aagaactcca aagtggactg caacggggct ggggccctgc acctcagcct
     3061 cagcttgctg ctgctcttct ctctcctcag cttattgtca ggtctggact acaaagacca
     3121 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agccgcggga
     3181 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc cagcccggga
     3241 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3421 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3661 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3721 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3781 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3841 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3961 cgagctgtac aagtaaagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     4021 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4081 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4141 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4201 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4261 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4321 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4381 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4441 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4501 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4561 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4621 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4681 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4741 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4861 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4981 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5041 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5101 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5161 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5221 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5281 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5341 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5401 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5461 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5521 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5581 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5641 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5701 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5761 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5821 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5881 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5941 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6001 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6061 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6121 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6181 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6241 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6301 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7081 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7201 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8401 atttccccga aaagtgccac ctgacgt
//
