LOCUS       V025925                 7644 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025925
VERSION     V025925
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..2324
                     /label="TTYH3(NM_025250)"
                     /note="TTYH3(NM_025250)"
                     /gene="TTYH3"
     CDS             2331..2396
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2420..3008
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3074..3122
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3324..3752
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3766..4095
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4162..4956
                     /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    5132..5253
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5302..5318)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5326..5342
                     /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(5350..5380)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5395..5416
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5704..6292)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6463..7323)
                     /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(7324..7428)
                     /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 ccaccatggc cggggtcagc tacgcggcgc
      781 cctggtgggt gagcctcctg caccggctgc cccacttcga cctgagctgg gaggccacta
      841 gcagccagtt ccggcccgag gacaccgact accagcaggc cctgctgctc ctgggggccg
      901 ccgccctggc ctgcctcgcc ctggacctcc tcttcctgct cttctactcc ttctggctgt
      961 gctgccggcg gcgcaagagc gaggagcacc tggacgccga ctgctgctgc acggcctggt
     1021 gtgtcatcat cgccacgctg gtgtgcagcg ccggcatcgc agtgggattc tacggcaacg
     1081 gggagaccag tgatggcatc catagggcca cctactcgct ccgccacgcc aaccgcacgg
     1141 tggccggggt ccaggaccgc gtgtgggaca cggcggtggg gctgaaccac acggcggagc
     1201 ccagcctgca gaccctggag cggcagctgg ccgggcggcc cgagcccctg cgagccgtac
     1261 agaggctgca gggcctgctg gagacgctgc tgggctacac ggccgccatc cccttttgga
     1321 ggaacacggc ggtgtcgctg gaggtgctgg cggagcaggt ggatctctac gactggtaca
     1381 ggtggctggg ctacctgggc ctgctgctgc tggacgtcat catctgcctc ctggtgctgg
     1441 ttggcctcat ccgcagctcc aagggcatcc tggtgggggt ctgcctgctg ggagtcctgg
     1501 ccctggtcat cagctggggc gcgctgggct tggagctggc tgtgtccgtg ggctccagcg
     1561 acttctgtgt ggaccctgac gcctacgtga ccaaaatggt ggaggagtac tcggtgctga
     1621 gtggggacat cctgcagtac tacctggcct gctcgccccg cgccgccaac cccttccagc
     1681 agaagctgtc gggcagccac aaggcactgg tggagatgca ggatgtcgtg gctgagcttc
     1741 tgaggaccgt cccctgggag cagccggcca ctaaggaccc cctcctccgc gtccaggagg
     1801 tgctgaatgg cacggaggtg aacctgcagc acctcaccgc cctggtggac tgccgcagcc
     1861 tgcatctgga ctacgtgcaa gcgctgaccg gcttctgcta tgacggcgtg gagggcctca
     1921 tctacctggc cctcttctcc ttcgtcacag ccctcatgtt cagctccatc gtctgcagcg
     1981 tcccgcacac ctggcagcaa aagagaggcc ctgatgagga cggggaggag gaggccgctc
     2041 cagggccgcg gcaggcgcac gacagcctct accgcgtcca catgcccagc ctgtacagct
     2101 gtggcagcag ctacggcagt gagaccagca tcccggccgc ggcccacacc gtcagcaacg
     2161 ccccggtcac tgagtacatg agccagaacg ctaatttcca gaacccccgc tgtgagaaca
     2221 ccccactcat tgggcgcgag tccccgccgc cctcatacac ctccagcatg agagccaaat
     2281 acctcgccac gagccagcct cgccctgact ccagcggcag ccacggatcc gactacaaag
     2341 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     2401 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2461 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2521 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2581 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2641 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2701 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2761 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2821 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2881 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2941 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3001 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3061 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     3121 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3181 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3241 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3301 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3361 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3421 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3481 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3541 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3601 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3661 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     3721 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3781 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3841 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3901 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3961 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4021 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4081 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4141 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4201 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4261 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4321 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4381 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4441 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4501 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4561 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4621 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4681 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4741 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4801 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4861 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4921 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     4981 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5041 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5101 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5161 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5221 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5281 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7441 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     7501 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7561 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7621 aattgcatga agaatctgct tagg
//
