LOCUS       V043992                 6730 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043992
VERSION     V043992
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..1386
                     /label="ZNRF1(NM_032268)"
                     /note="ZNRF1(NM_032268)"
                     /gene="ZNRF1"
     CDS             1393..1482
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1506..2094
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2160..2208
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2410..2838
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2852..3181
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3248..4042
                     /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    4218..4339
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4388..4404)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4412..4428
                     /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(4436..4466)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4481..4502
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4790..5378)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5549..6409)
                     /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(6410..6514)
                     /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 ccaccatggg gggcaagcag
      721 agcacggcgg cccgctcccg gggccccttc ccgggggtct ccaccgatga cagcgccgtg
      781 ccgccgccgg gaggggcgcc ccatttcggg cactaccgga cgggcggcgg ggccatgggg
      841 ctgcgcagcc gctcggtcag ctcggtggca ggcatgggca tggaccccag cacggccggg
      901 ggggtgccct ttggcctcta cacccccgcc tcccggggca ccggcgactc cgagagggcg
      961 cccggcggcg gagggtctgc gtccgactcc acctatgccc atggcaatgg ttaccaggag
     1021 acgggcggcg gtcaccatag agacgggatg ctgtacctgg gctcccgagc ctcgctggcg
     1081 gatgctctac ctctgcacat cgcacccagg tggttcagct cgcatagtgg tttcaagtgc
     1141 cccatttgct ccaagtctgt ggcttctgac gagatggaaa tgcactttat aatgtgtttg
     1201 agcaaacctc gcctctccta caacgatgat gtgctgacta aagacgcggg tgagtgtgtg
     1261 atctgcctgg aggagctgct gcagggggac acgatagcca ggctgccctg cctgtgcatc
     1321 tatcacaaaa gctgcataga ctcgtggttt gaagtgaaca gatcttgtcc ggaacaccct
     1381 gcggacctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     1441 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc
     1501 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1561 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1621 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1681 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     1741 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     1801 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     1861 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     1921 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     1981 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2041 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     2101 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     2161 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     2221 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     2281 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     2341 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     2401 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2461 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2521 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     2581 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2641 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2701 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2761 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     2821 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3061 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3121 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3181 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3241 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3301 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3361 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3421 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3481 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3541 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3601 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3661 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3721 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3781 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3841 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3901 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3961 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4021 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     4081 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4141 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4201 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4321 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4381 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4441 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4501 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4561 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4621 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4681 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4741 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4981 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5161 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5221 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5281 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5341 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5401 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5461 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5521 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5581 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5641 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5701 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5761 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5821 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5881 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5941 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6001 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6061 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6121 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6181 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6241 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6301 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6361 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6421 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6481 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6541 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     6601 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6661 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6721 tctgcttagg
//
