LOCUS       V023196                 7854 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023196
VERSION     V023196
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..2528
                     /label="Pdlim5(NM_019808)"
                     /note="Pdlim5(NM_019808)"
                     /gene="Pdlim5"
     CDS             2541..2606
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2630..3218
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3284..3332
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3534..3962
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3976..4305
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4372..5166
                     /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    5342..5463
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5512..5528)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5536..5552
                     /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(5560..5590)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5605..5626
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5914..6502)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6673..7533)
                     /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(7534..7638)
                     /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 ctcggatccg ccaccatgag caactacagt gtgtcattgg
      781 tcggcccagc tccttggggt ttccggctcc aaggtggcaa ggatttcaac atgcctctga
      841 caatctctag tctgaaggat ggtggcaagg catctcaggc acatgtcaga ataggggacg
      901 tggttctcag catcgatggg atcagtgcac agggaatgac gcatcttgaa gcccagaaca
      961 agattaaggc ttgtacgggc tccttgaata tgactctaca aagagcttca gctgcagcca
     1021 agagtgagcc ggtttccgtc cagaagggtg aacctaagga agtagttaag cctgtgccca
     1081 ttacatctcc tgctgtatcc aaagtcactt ccactaccaa catggcctac aataaagcac
     1141 cacggccctt tggttctgtg tcttcaccaa aagtcacatc catcccatca ccatcgtctg
     1201 cgttcacccc agcccatgcc gccacttcat cacatgcttc ccccacacct gtggccgctg
     1261 ccactcccct ccacctctct gcatccggac tgcatgttag tgccaatctt agtgctgacc
     1321 agtgttcatc tccaccgaac actggtaaac ctgcagttaa tgtcccacgg cagcccacag
     1381 tcaccagcgt gtgttccgag tctgctcagg agctagcaga gggacagaga agaggatccc
     1441 agggtgacat taagcagcaa aatggcccac caagaaaaca cattgtggag cgcaacacgg
     1501 agttttatca catacccact cacagtgatg ccagcaagaa acggctgatt gaggacactg
     1561 aggactggcg cccccggact ggaacgactc agtctcgttc tttccggatc cttgcccaga
     1621 tcactgggac tgagcatctg acagaatctg aaaatgacaa tacgaagaag gcaaatagca
     1681 cccaggagcc ttctcagcaa ccggcgtcat ctggagcttc acctctgagc gcgtctgagg
     1741 gccccgagag cccaggttcc agcaggccct cggtggctgg cctccgctcc gctgctgctt
     1801 tcaagcctgt aggatccacc agcgtcaagt cacctagctg gcagcgtcca aaccaagcag
     1861 caccttctac tgggagaatt tcaaacaatg cacgttcttc aggtacaggg gcgtctgtgg
     1921 ggcctcccca gccaagtgac caagacacac ttgtgcagag agcggaacac attccagcgg
     1981 gcaagcggac ccccatgtgt gcccactgca accaagttat caggggacct tttcttgtgg
     2041 ctctggggaa gtcctggcat ccagaagaat ttaactgtgc tcattgcaaa aacacaatgg
     2101 cctacatagg ctttgtagag gagaagggag cactgtactg tgagctatgc tatgagaagt
     2161 tctttgctcc tgaatgtggc cggtgccaaa ggaagatcct tggagaagtc atcaatgctt
     2221 tgaagcagac ctggcatgtg tcctgttttg tgtgtgtggc ctgtggaaaa cccattcgta
     2281 ataatgtttt ccacttggaa gatggcgagc cctactgtga gacagactac tacgcccttt
     2341 ttggaacaat atgtcgtgga tgtgaattcc ccatagaggc tggtgacatg ttcctggaag
     2401 ctctgggata cacttggcat gatacttgct ttgtgtgctc tgtgtgttgt gaaagtttgg
     2461 aagggcagac atttttctcc aagaaggaca agccactttg caaaaaacat gctcattctg
     2521 tgaattttct cgagtctaga gactacaaag accatgacgg tgattataaa gatcatgaca
     2581 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     2641 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2701 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2761 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2821 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2881 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2941 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3001 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3061 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3121 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3181 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3241 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3301 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3361 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3421 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3481 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3541 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3601 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3661 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3721 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3781 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3841 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3901 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3961 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4321 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4381 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4441 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4501 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4561 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4621 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4681 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4741 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4801 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4861 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4921 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4981 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5041 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5101 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5161 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5221 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5281 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5341 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5401 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5461 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5521 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5581 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5641 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5701 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5761 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5821 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5941 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6541 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6601 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6661 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6721 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6781 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6841 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6901 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6961 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7021 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7081 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7141 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7201 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7261 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7321 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7381 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7441 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7501 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7561 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7621 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7681 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7741 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7801 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
