LOCUS       V034826                 9177 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034826
VERSION     V034826
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..3857
                     /label="TSHZ2(NM_173485)"
                     /note="TSHZ2(NM_173485)"
                     /gene="TSHZ2"
     CDS             3864..3929
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3953..4541
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4607..4655
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4857..5285
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5299..5628
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5695..6489
                     /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    6665..6786
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6835..6851)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6859..6875
                     /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(6883..6913)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6928..6949
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7237..7825)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7996..8856)
                     /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(8857..8961)
                     /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 ccaccatgcc gaggagaaaa cagcaggcac
      781 ccaagcgggc ggcaggctac gcccaggagg aacagctgaa agaagaggag gaaataaaag
      841 aagaggagga ggaggaggac agcggttcag tagctcaact gcagggtggc aatgacacag
      901 ggacggacga ggagctagaa acgggcccag agcaaaaagg ctgcttcagc taccagaact
      961 ctccaggaag tcatttgtcc aatcaggatg ccgagaacga gtctctgctg agtgacgcca
     1021 gtgatcaggt gtcggacatc aagagtgtct gcggcagaga tgcctcagac aagaaagcac
     1081 acactcacgt caggcttcca aacgaagcac acaattgcat ggataaaatg accgctgtct
     1141 acgccaacat cctgtcggat tcctactggt caggcctggg ccttggcttc aagctgtcca
     1201 atagtgagag gaggaactgt gacacccgaa acggcagcaa caagagtgat tttgattggc
     1261 accaagacgc tctgtccaaa agcctgcagc agaacttgcc ttctcggtcc gtctcgaaac
     1321 ccagcctgtt cagctcggtg cagttgtacc gacagagcag caagatgtgc gggactgtgt
     1381 tcacaggggc cagcagattc cgatgccgac agtgcagcgc ggcctatgac accctagtcg
     1441 agctgactgt gcacatgaat gaaacgggcc actatcaaga tgacaaccgc aaaaaggaca
     1501 agctcagacc cacgagctat tcaaagccca ggaaaagggc tttccaggat atggacaaag
     1561 aggatgctca aaaggttctg aaatgtatgt tttgtggcga ctcctttgat tccctccaag
     1621 atttgagcgt ccacatgatt aaaacaaaac attaccaaaa agtgcctttg aaggagccag
     1681 tcccaaccat ttcctcgaaa atggtcaccc cggctaagaa acgcgttttt gatgtcaatc
     1741 ggccgtgttc ccccgattca accacaggat cttttgcaga ttctttttct tctcagaaga
     1801 acgccaactt gcagttgtcc tccaacaacc gctatggcta ccaaaatgga gccagctaca
     1861 cctggcagtt tgaggcctgc aagtcccaga tcttaaagtg catggagtgt gggagctccc
     1921 atgacacctt gcagcagctc accacccaca tgatggtcac aggtcacttt ctcaaggtca
     1981 ccagctctgc ctccaagaaa gggaagcagc tggtattaga cccgttagca gtggagaaaa
     2041 tgcagtcgtt gtctgaggcc ccaaacagtg attctctggc tcccaagcca tccagtaact
     2101 cagcatcaga ttgtacagcc tctacaactg agttaaagaa agagagtaaa aaagaaaggc
     2161 cagaggaaac cagcaaggat gagaaagtcg tgaaaagcga ggactatgaa gatcctctac
     2221 aaaaaccttt agaccctaca atcaaatatc aatacctaag ggaggaagac ttggaagatg
     2281 gctcaaaggg tggaggggac attttgaaat ctttggaaaa tactgtcacc acagccatca
     2341 acaaagccca aaacggggcc cccagctgga gtgcctaccc cagcatccac gcagcctacc
     2401 agctgtctga gggcaccaag ccgcctttgc ctatgggatc ccaggtactg cagatccggc
     2461 ctaatctcac caacaagctg aggcccattg caccaaagtg gaaagtgatg ccactggttt
     2521 ctatgcccac acacctggcc ccttacactc aagtcaagaa agagtcagaa gacaaagatg
     2581 aagcggtgaa ggagtgtggg aaagaaagtc cccacgaaga ggcctcatct ttcagccaca
     2641 gtgagggcga ttctttccgc aaaagtgaaa cacctccaga agccaaaaag accgagctgg
     2701 gtcccctgaa ggaggaggag aagctgatga aagagggcag cgagaaggag aaaccccagc
     2761 ccctggagcc cacatctgct ctgagcaatg ggtgcgccct cgccaaccac gccccggccc
     2821 tgccatgcat caacccactc agcgccctgc agtccgtcct gaacaatcac ttgggcaaag
     2881 ccacggagcc cttgcgctca ccttcctgct ccagcccaag ttcaagcaca atttccatgt
     2941 tccacaagtc gaatctcaat gtcatggaca agccggtctt gagtcctgcc tccacaaggt
     3001 cagccagcgt gtccaggcgc tacctgtttg agaacagcga tcagcccatt gacctgacca
     3061 agtccaaaag caagaaagcc gagtcctcgc aagcacaatc ttgtatgtcc ccacctcaga
     3121 agcacgctct gtctgacatc gccgacatgg tcaaagtcct ccccaaagcc accaccccaa
     3181 agccagcctc ctcctccagg gtccccccca tgaagctgga aatggatgtc aggcgctttg
     3241 aggatgtctc cagtgaagtc tcaactttgc ataaaagaaa aggccggcag tccaactgga
     3301 atcctcagca tcttctgatt ctacaagccc agtttgcctc gagcctcttc cagacatcag
     3361 agggcaaata cctgctgtct gatctgggcc cacaagagcg tatgcaaatc tctaagttta
     3421 cgggactctc aatgaccact atcagtcact ggctggccaa cgtcaagtac cagcttagga
     3481 aaacgggcgg gacaaaattt ctgaaaaaca tggacaaagg ccaccccatc ttttattgca
     3541 gtgactgtgc ctcccagttc agaacccctt ctacctacat cagtcactta gaatctcacc
     3601 tgggtttcca aatgaaggac atgacccgct tgtcagtgga ccagcaaagc aaggtggagc
     3661 aagagatctc ccgggtatcg tcggctcaga ggtctccaga aacaatagct gccgaagagg
     3721 acacagactc taaattcaag tgtaagttgt gctgtcggac atttgtgagc aaacatgcgg
     3781 taaaactcca cctaagcaaa acgcacagca agtcacccga acaccattca cagtttgtaa
     3841 cagacgtgga tgaagaagga tccgactaca aagaccatga cggtgattat aaagatcatg
     3901 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     3961 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4021 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4081 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4141 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4201 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4261 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4321 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4381 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4441 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4501 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     4561 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4621 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4681 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4741 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4801 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     4861 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4921 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4981 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     5041 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5101 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5161 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5221 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5281 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5401 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5461 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5581 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5641 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5701 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5761 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5821 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5881 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5941 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6001 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6061 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6121 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6181 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6241 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6301 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6361 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6421 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6481 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     6541 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6661 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6781 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     9001 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     9061 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     9121 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
