LOCUS       V019523                 8186 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019523
VERSION     V019523
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             683..2866
                     /label="PCCA(NM_000282)"
                     /note="PCCA(NM_000282)"
                     /gene="PCCA"
     CDS             2873..2938
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2962..3550
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3616..3664
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3866..4294
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4308..4637
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4704..5498
                     /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    5674..5795
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5844..5860)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5868..5884
                     /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(5892..5922)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5937..5958
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6246..6834)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7005..7865)
                     /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(7866..7970)
                     /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 ggatccgcca ccatggcggg gttctgggtc gggacagcac cgctggtcgc
      721 tgccggacgg cgtgggcggt ggccgccgca gcagctgatg ctgagcgcgg cgctgcggac
      781 cctgaagcat gttctgtact attcaagaca gtgcttaatg gtgtcccgta atcttggttc
      841 agtgggatat gatcctaatg aaaaaacttt tgataaaatt cttgttgcta atagaggaga
      901 aattgcatgt cgggttatta gaacttgcaa gaagatgggc attaagacag ttgccatcca
      961 cagtgatgtt gatgctagtt ctgttcatgt gaaaatggcg gatgaggctg tctgtgttgg
     1021 cccagctccc accagtaaaa gctacctcaa catggatgcc atcatggaag ccattaagaa
     1081 aaccagggcc caagctgtac atccaggtta tggattcctt tcagaaaaca aagaatttgc
     1141 cagatgtttg gcagcagaag atgtcgtttt cattggacct gacacacatg ctattcaagc
     1201 catgggcgac aagattgaaa gcaaattatt agctaagaaa gcagaggtta atacaatccc
     1261 tggctttgat ggagtagtca aggatgcaga agaagctgtc agaattgcaa gggaaattgg
     1321 ctaccctgtc atgatcaagg cctcagcagg tggtggtggg aaaggcatgc gcattgcttg
     1381 ggatgatgaa gagaccaggg atggttttag attgtcatct caagaagctg cttctagttt
     1441 tggcgatgat agactactaa tagaaaaatt tattgataat cctcgtcata tagaaatcca
     1501 ggttctaggt gataaacatg ggaatgcttt atggcttaat gaaagagagt gctcaattca
     1561 gagaagaaat cagaaggtgg tggaggaagc accaagcatt tttttggatg cggagactcg
     1621 aagagcgatg ggagaacaag ctgtagctct tgccagagca gtaaaatatt cctctgctgg
     1681 gaccgtggag ttccttgtgg actctaagaa gaatttttat ttcttggaaa tgaatacaag
     1741 actccaggtt gagcatcctg tcacagaatg cattactggc ctggacctag tccaggaaat
     1801 gatccgtgtt gctaagggct accctctcag gcacaaacaa gctgatattc gcatcaacgg
     1861 ctgggcagtt gaatgtcggg tttatgctga ggacccctac aagtcttttg gtttaccatc
     1921 tattgggaga ttgtctcagt accaagaacc gttacatcta cctggtgtcc gagtggacag
     1981 tggcatccaa ccaggaagtg atattagcat ttattatgat cctatgattt caaaactaat
     2041 cacatatggc tctgatagaa ctgaggcact gaagagaatg gcagatgcac tggataacta
     2101 tgttattcga ggtgttacac ataatattgc attacttcga gaggtgataa tcaactcacg
     2161 ctttgtaaaa ggagacatca gcactaaatt tctctccgat gtgtatcctg atggcttcaa
     2221 aggacacatg ctaaccaaga gtgagaagaa ccagttattg gcaatagcat catcattgtt
     2281 tgtggcattc cagttaagag cacaacattt tcaagaaaat tcaagaatgc ctgttattaa
     2341 accagacata gccaactggg agctctcagt aaaattgcat gataaagttc ataccgtagt
     2401 agcatcaaac aatgggtcag tgttctcggt ggaagttgat gggtcgaaac taaatgtgac
     2461 cagcacgtgg aacctggctt cgcccttatt gtctgtcagc gttgatggca ctcagaggac
     2521 tgtccagtgt ctttctcgag aagcaggtgg aaacatgagc attcagtttc ttggtacagt
     2581 gtacaaggtg aatatcttaa ccagacttgc cgcagaattg aacaaattta tgctggaaaa
     2641 agtgactgag gacacaagca gtgttctgcg ttccccgatg cccggagtgg tggtggccgt
     2701 ctctgtcaag cctggagacg cggtagcaga aggtcaagaa atttgtgtga ttgaagccat
     2761 gaaaatgcag aatagtatga cagctgggaa aactggcacg gtgaaatctg tgcactgtca
     2821 agctggagac acagttggag aaggggatct gctcgtggag ctggaactcg aggactacaa
     2881 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     2941 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     3001 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3061 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3121 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3181 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3241 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3301 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3361 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3421 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3481 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3541 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3601 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     3661 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     3721 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     3781 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     3841 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3901 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3961 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4021 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4081 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4141 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4201 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     4261 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4441 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4561 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4621 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4681 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4741 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4801 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4861 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4921 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4981 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5041 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5101 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5161 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5221 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5281 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5341 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5401 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5461 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     5521 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5581 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5641 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5701 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5761 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5821 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5881 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5941 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6001 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6061 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6121 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6181 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6241 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6301 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6361 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6421 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6481 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6541 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6601 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6661 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6721 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6781 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6841 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6901 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6961 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7021 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7081 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7141 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7201 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7261 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7321 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7381 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7441 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7501 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7561 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7621 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7681 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7741 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7801 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7861 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7921 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7981 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     8041 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     8101 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8161 acaattgcat gaagaatctg cttagg
//
