LOCUS       V047080                 7149 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047080
VERSION     V047080
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1829
                     /label="Gja5(NM_001271628)"
                     /note="Gja5(NM_001271628)"
                     /gene="Gja5"
     CDS             1836..1901
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1925..2513
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2579..2627
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2829..3257
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3271..3600
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3667..4461
                     /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    4637..4758
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4807..4823)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4831..4847
                     /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(4855..4885)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4900..4921
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5209..5797)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5968..6828)
                     /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(6829..6933)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg tgactggagc ttcctggggg
      781 agttcctgga ggaggtccac aagcactcca cagtcatcgg caaggtctgg ctcactgtcc
      841 tgttcatttt ccgcatgctg gtcctgggca ccgctgctga gtcctcctgg ggagatgagc
      901 aggccgactt ccggtgcgat accattcagc ctggttgcca aaatgtctgc tatgaccaag
      961 ccttccccat ctcccacatt cgttattggg tactgcagat catctttgtg tccacgcctt
     1021 ctctagtgta catgggccat gccatgcaca ctgtgcgcat gcaggaaaag cagaaattgc
     1081 gggatgctga gaaagctaaa gaggcccacc gcactggtgc ctatgagtac ccagtagccg
     1141 aaaaggccga gctgtcctgc tggaaagaag tagatgggaa gattgtcctc cagggcaccc
     1201 tactcaacac ctatgtctgc accattctga tccgcaccac catggaggtg gccttcatcg
     1261 taggccagta cctcctctat gggatcttcc tggataccct gcatgtctgc cgcaggagtc
     1321 cctgtcccca cccagtcaac tgttatgttt cgaggcccac ggagaagaat gtcttcattg
     1381 tctttatgat ggctgtggct ggactgtctc tgtttctcag cctggctgaa ctctaccacc
     1441 tgggctggaa gaagatccga cagcgctttg gcaagtcacg gcagggtgtg gacaagcacc
     1501 agctgcctgg ccctcccacc agcctcgtcc agagcctcac tcctccccct gacttcaatc
     1561 agtgcctaaa gaacagctcc ggagagaaat tcttcagcga cttcagtaat aacatgggct
     1621 cccggaagaa tccagacgct ctggccactg gggaagtgcc aaaccaggag cagattccag
     1681 gggaaggctt catccacatg cactatagcc agaagccaga gtacgccagt ggagcctctg
     1741 cgggccaccg ccttcctcag ggctaccata gtgacaaacg gcgccttagt aaggccagca
     1801 gcaaagcaag gtcagatgac ctgtcagtgg gatccgacta caaagaccat gacggtgatt
     1861 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     1921 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1981 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2041 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2101 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2161 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2221 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2281 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2341 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2401 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2461 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2521 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2581 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     2641 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     2701 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     2761 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     2821 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2881 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2941 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3001 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3061 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3121 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3181 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3241 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3361 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3421 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3481 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3541 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3601 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3661 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3721 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3781 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3841 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3901 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3961 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4021 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4081 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4141 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4201 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4261 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4321 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4381 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4441 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4501 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4561 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4621 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4741 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4801 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4861 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4921 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4981 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5401 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5641 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5941 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6001 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6061 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6121 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6181 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6241 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6301 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6361 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6421 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6481 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6541 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6601 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6661 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6721 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6781 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6841 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6901 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6961 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7021 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7081 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7141 ctgcttagg
//
