LOCUS       V043256                 6646 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043256
VERSION     V043256
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             964..2136
                     /label="Dap3(NM_001368419)"
                     /note="Dap3(NM_001368419)"
                     /gene="Dap3"
     CDS             2143..2208
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2246..2470
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2516..2944
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2958..3287
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3354..4148
                     /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    4322..4443
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4492..4508)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4516..4532
                     /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(4540..4570)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4585..4606
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4894..5479)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5650..6510)
                     /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(6511..6615)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgctga caggaataac aaggcttttc tccagggtcc agaagttgga cccgaggtgt
     1021 tttttgcaca tgagtgttca agcgactcag aacagtcagg ttccagctga gcgtcccaga
     1081 actgtttccc gcaccagtga cagtgacccg gctaagcatg gggagcagca tgagggtcag
     1141 cactacagca tccccctcca ggatctgaag acagtgtttc ctcacggcct gcctcctcgg
     1201 tacatgatgc aggtgaagac ttttggcgaa gcttgtctga tggtcaggaa gccagctctc
     1261 gagcttctgg gctacctgaa aaataccaat tttgctcatc cagctgtgcg atatcttctt
     1321 tatggggaga aggggacagg aaagaccctc agtctctgcc atgctgttca tttttgtgca
     1381 agacatgact ggctgattct gcatatccca gatgctcatc tttgggtaaa aaactgccgg
     1441 gagcttctac agtccactca caacaaacag cgctttgatc aacccttaga ggcctctacc
     1501 tggctgaaga atttcaaaac taccaatgag cgcttcctga gtcagataaa agttcaagag
     1561 aagtatgttt ggaataagag agaaagcact gagaaaggca gtcctctggg agaagttgtt
     1621 gaacagggcc taacccgagt gaggaatgcc actgacgctg ttggggtcgt gctgaaggag
     1681 ctgaaggctc agagcgccct agggctcttt cacctcctgg tggctgtgga tggagtcaat
     1741 gccctctggg gaaggaccac actgaaaaaa gaagatagaa ccttgattgc cccagaggaa
     1801 ctctcccttg tccacaatct aaggaaaatg gtgaaaaatg actggcatgg aggtgcaatc
     1861 gtgttgagct tgagccaaac cgggtctctc tttaagtcca gaacagccta tttgccgcat
     1921 gagctgctgg gaaaggaagg atttaatgcc ctggagcctt ttcttcccat cctcattccc
     1981 aactataatc caaaggagtt tgaaagttcc ttccagtact acttagagaa caactggctt
     2041 caacatgaga aagcttcaac agaagaaggg aggaaggagc tgaggttcct gagtaactgc
     2101 aaccctgagc agctggagcg gctctgtgcc tcgctgctcg aggactacaa agaccatgac
     2161 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     2221 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2401 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2461 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2941 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3301 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3361 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3421 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3481 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3541 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3601 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3661 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3721 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3781 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3841 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3901 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3961 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4021 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4081 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4141 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4201 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4261 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4321 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4381 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4441 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4501 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4561 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4621 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4681 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4741 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4801 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4861 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4921 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4981 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5041 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5101 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5161 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5221 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5281 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5341 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5401 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5701 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
