LOCUS       V037200                 9654 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037200
VERSION     V037200
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..4334
                     /label="Ttll4(NM_001014974)"
                     /note="Ttll4(NM_001014974)"
                     /gene="Ttll4"
     CDS             4341..4406
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4430..5018
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5084..5132
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5334..5762
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5776..6105
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6172..6966
                     /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    7142..7263
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7312..7328)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7336..7352
                     /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(7360..7390)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7405..7426
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7714..8302)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8473..9333)
                     /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(9334..9438)
                     /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 ctcagcagga acagagcact
      781 acagtattgg cctgcgccga ggaaacagct tcaagcagag acatccctca ggcacagtgt
      841 ctgcttcacc atctgagaaa ccctcggagg tcaaagtctg gtctcaggcc catcagcagg
      901 tgaagccaat ctggaaactg gaaaagaagc acgtaggcac actgtcagca gggctgggca
      961 ccagtttctt gggagtccca tcccagccag cctatttcct ttgccctagc actttatgta
     1021 gctctgggac cacagcagtc attgcaggcc acagcaaccc ctgttacctg cagtccctcc
     1081 ctaacttgtt cagcaatact ctgctgtacc gccgcaccaa cgtgaggcaa aaaccatacc
     1141 agcaactgga atctttctgc ttgcgttcca gcccttcaga gaaaagatct ttttctctcc
     1201 ctcagaaggg tcttcctgtc agtgtcactg ccaataaagc cacatcttct acagtcttcc
     1261 ccatggccca gcccatggca acttcaccca ctgatccata cctctcactg gcagcagctg
     1321 gggagaaccc ttcaaggaag agcctggcct ctgctatctc agggaagatt gcatctccac
     1381 tgtcctataa gcccatgctg aacaacaact ccttcatgag gccaaatagc actaaagtgc
     1441 ccttatcaca agccacagat ggcctgaagc cagtatcctc acccaagatc cagcctgtct
     1501 catggcatca ttcagggggt actggggact gtgtacccca acctggtgac cataaggtgc
     1561 cccaaaacat tgctactgtc ttagatgatg tcactgcccc tatcacccca tctatcccta
     1621 gcaccctcaa catatccaca gccagtgtta cctcttccca gtgcagtcag agtaacttta
     1681 ggatggaggc acatccctgt ggcctggatg aaaacccgga ttcccagagt gctactaaag
     1741 aggtacactt cactgaggct gtgaggaagt tggctgaaaa aggcttggag aagatgccac
     1801 gacaaggcta ccagtttgaa caggcttgtt ttgtgaaccc cagcttccag tggggtcttc
     1861 tcaacagaag caggcggtgg aaacctctga tgggccagcg gtttcctcag gaggatattg
     1921 gattagacag tgcgatcctt cctggtactt cagacacctt ggggttggac agcacagtct
     1981 tctgtaccaa gaggatcagc attcacctcc ttgcctcaca tgtccatggg ctcaatccca
     2041 gccctgcctg tgggtctgca gttgaccccc aggtacttgg agaagacaga gctcctgttc
     2101 ccccttcttc cctccagcct cttggtgtag ctgaagtggc cactcgtctt tcttctgtcc
     2161 atcttgacca gcctgggaag gagcctgagg aagccaagga cctgaactct tgtactaagg
     2221 gtggtggttc agctactgac ctccagccaa accaggtgga gcctgaagat acagaagatg
     2281 aactgggaga tggtttggaa gatagctgca gccatgatga gaatgaagaa gaggaaggag
     2341 actcagagtg ctcttcttta agtgttgtct cccccagcga gtcagtggca ttaatatctc
     2401 ggaactgtgt ggatttgatg tccaaatccc ttcccaatca tgagaaagtt gtacggccag
     2461 ccctcatcta cagtctcttc cccaatgtaa cccctaccat ctattttggg actcgggatg
     2521 aaagagtgga gaaacttcct tgggaacaga ggaggctgct tcggtggaag atgagcacag
     2581 tgacccccaa cattgtcaag cagaccatcg gtcggtcaca cttcaaaatc agcaagagaa
     2641 atgatgactg gctgggctgc tggggtcacc acatgaagtc tcctggtttt cgatccatta
     2701 gagagcatca gaagctaaac cacttcccag gctccttcca gattgggagg aaggaccggc
     2761 tgtggcggaa cctgtctcgc atgcaaagcc gttttggcaa gaaggagttc agtttctttc
     2821 ctcagtcctt tatcctgccc caggattcca agctcttgcg caaagcctgg gagagcagca
     2881 gccgccagaa gtggattgtg aagccaccag catctgctcg aggcattggc attcaggtca
     2941 ttcacaaatg gagtcagctc cccaagcgca ggcccctcct ggtacagagg tatctacaca
     3001 agccctacct cattagtggc agcaaatttg atcttcggat ctatgtttac gtgacttctt
     3061 atgatcctct acggatttac ctcttttcgg atggactcgt ccgttttgcc agttgcaagt
     3121 attccccttc catgaagagt cttagcaaca agttcatgca cctgaccaac tacagtgtca
     3181 ataaaaagaa cacagaatac caggctaatg ctgatgagac tgcatgccaa gggcacaaat
     3241 gggcattgaa agctttgtgg aactatctga gccagaaagg aattaatagt gatgccatct
     3301 gggagaagat aaaggatgtt gtagtcaaaa ccatcatctc gtcagagccc tatgtgacta
     3361 acctactgaa gttatatgtg cggcgcccct atagctgtca tgagctcttt ggttttgaca
     3421 tcatgctgga tgagaacctg aaaccctggg tcctagaagt caacatttct ccaagcctcc
     3481 attccaactc tcctctggac atcagcatca aaggccagat gatccgagac cttctgaacc
     3541 tggctggctt tgttctgccc aacatggagg atatcatctc tagttccagc agccccagca
     3601 gctccagcgg ctccagcacc agcttgccaa gctctcctag agacaaatgt caaatgactc
     3661 cagagcactt cactgcacag aagatgaaga aagcctatta cttgacccag aaaattcctg
     3721 accaggactt ctatgcgtcc gtgctggacg tcttgacacc agatgatgtt cgggttttgg
     3781 tggagatgga agatgagttt tctcgtcgtg gtcaatttga aagaattttt ccttcccgaa
     3841 tctcttctcg atatcttcgg ttttttgagc agccacgata tttcaacatt ctcaccaccc
     3901 agtgggaaca gaagtaccat ggaaacaagc tcaaaggagt agatctgctt cggaattggt
     3961 gctacaaagg cttccacacg gggattgtct ctgactcagc cccactgtgg tctctgccaa
     4021 catcgctcat gactacctca aagggtgatg ggacaccaaa ttccgccagc aagtcacgaa
     4081 agaagagcgc ctctgaggga accacactgt cctctgaaga caggtcaaca cccaagtcta
     4141 agaagagtca agctggcctg tctcccatat ccaggaagac tttgtcctca aggagcaatg
     4201 agaacaccag caaacagtcc aagcgctcta ccccggggtt acctgtcctc aagtactctg
     4261 ggcagagttc cagactttct gctgcttctg cttcccagtc agtcactgac tcccgcttga
     4321 ctgctgtgag ctcgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     4381 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     4441 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4501 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4561 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4621 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4681 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4741 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4801 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     4861 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4921 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4981 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     5041 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5101 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5161 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5221 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5281 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     5341 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5401 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5461 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     5521 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5581 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5641 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5701 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5761 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     6061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     6121 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     6181 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     6241 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6301 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     6361 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6421 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6481 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     6541 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     6601 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     6661 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     6721 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     6781 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     6841 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6901 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6961 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     7021 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7081 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7141 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7261 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7321 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7381 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7501 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7561 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7621 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     7921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     8161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8521 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8581 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8641 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8701 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8761 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8821 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8881 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8941 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9001 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9061 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9121 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9181 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9241 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9301 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9361 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9421 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     9481 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     9541 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     9601 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
