LOCUS       V023322                 8867 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023322
VERSION     V023322
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             755..3541
                     /label="TEX10(NM_017746)"
                     /note="TEX10(NM_017746)"
                     /gene="TEX10"
     CDS             3554..3619
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3643..4231
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4297..4345
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4547..4975
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4989..5318
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5385..6179
                     /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    6355..6476
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6525..6541)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6549..6565
                     /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(6573..6603)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6618..6639
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6927..7515)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7686..8546)
                     /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(8547..8651)
                     /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 ctcggtccgc caccatgact aaaaaaagaa aacgccaaca
      781 tgattttcaa aaagtaaaat tgaaagttgg taaaaagaag cccaagttac aaaatgctac
      841 tcctacaaac tttaaaacaa agactataca tctgcctgag caactcaaag aggatggaac
      901 acttccaaca aacaatagaa aacttaacat aaaggatttg ctgtcacaga tgcatcacta
      961 caatgctggg gttaaacaaa gtgctcttct tggacttaaa gaccttttgt ctcaataccc
     1021 atttataatt gatgcacacc tttcaaacat attaagtgaa gtgactgctg tgtttacaga
     1081 taaagatgct aatgtacgat tagcagcagt tcaacttctt caattcctgg cccccaaaat
     1141 acgagctgaa caaatttctc cattttttcc tttggtaagt gcccatctct ctagtgccat
     1201 gactcacatt actgaaggaa ttcaggagga ctctttaaaa gttttggaca ttctgctgga
     1261 acagtaccca gctctaatta ctggccgtag cagcatattg cttaagaatt ttgtagaact
     1321 tatttctcat cagcagctgt ccaaaggact gataaataga gacagatccc agtcctggat
     1381 actttctgta aatcctaatc ggagactcac ttctcagcaa tggaggctga aagtcttagt
     1441 gagactcagt aaattccttc aggccttggc agatggatcc agtaggttga gagaaagtga
     1501 aggacttcag gaacagaaag aaaatcccca tgccactagc aactccattt ttatcaactg
     1561 gaaggaacat gccaacgacc agcaacacat ccaggtttat gaaaatgggg gttcacagcc
     1621 aaatgtcagt tcacagttca ggctacggta tctggttgga ggactgagtg gtgtggatga
     1681 aggcctgtca tctactgaaa acctgaaagg atttattgag ataataattc cattgctaat
     1741 tgaatgctgg gttgaagctg tacctccaca actagctact cctgttggga atggtataga
     1801 acgagaacct ctacaggtta tgcagcaagt tcttaatatt atttcccttc tgtggaaact
     1861 ctctaaacaa caggatgaaa cccataaatt ggagtcatgg cttcgaaaga actaccttat
     1921 tgattttaaa caccatttta tgagtcgttt tccatatgtc ttaaaagaaa taaccaagca
     1981 caaaaggaaa gagccaaata aaagcatcaa gcattgcaca gttctctcca ataacataga
     2041 tcatctctta ctgaatttaa cactgtctga tatcatggtc tccctggcaa atgcgtcaac
     2101 tttgcagaag gattgcagtt ggatagaaat gataaggaaa tttgtaacag agacccttga
     2161 agatggctct aggctaaata gtaagcaact gaacagattg ctgggagtat cctggaggtt
     2221 aatgcaaata cagccaaaca gagaggacac agagactctt attaaggcag tttatacatt
     2281 atatcagcag aggggcctta tccttccagt tcggactttg ttattgaagt ttttcagtaa
     2341 aatctatcag acagaagaac tgagatcttg tagattcaga tatcgtagta aagtgttatc
     2401 ccgttggctg gctggcttac cattgcaact tgctcatctt ggctcccgaa atcctgagct
     2461 ctctacacag cttatcgata tcattcatac cgctgcagca cgagcaaata aagaattact
     2521 aaaaagttta caagctactg ccctccgaat ttatgatcca caagaaggtg ctgtggtggt
     2581 tctccctgca gactctcagc agcgtttggt tcagcttgta tatttcctac ccagtctgcc
     2641 ggctgatttg ctttctcggt taagtcgttg ctgtattatg ggaagactca gttcaagttt
     2701 ggctgccatg cttatcggga tactgcacat gagatcatca ttttctgggt ggaagtattc
     2761 agctaaagac tggttgatga gtgatgtaga ctatttcagc ttcttatttt ccacacttac
     2821 agggttttcg aaagaggagt tgacttggct tcagagcctt cgaggagttc ctcatgtcat
     2881 ccagacacag ctttcccctg tgcttctcta ccttacagat ttggatcaat ttttacacca
     2941 ctgggatgta acagaggcag tttttcacag tttattggtt attcctgccc gaagtcagaa
     3001 ctttgacatc ttgcaaagtg ccatcagtaa gcatttggtt gggttgactg taattcctga
     3061 cagcacggct ggctgtgttt ttggtgttat ctgtaagctc ctggatcata cttgtgtagt
     3121 tagtgagact ctactgccat ttctggcttc ttgttgctac agtcttcttt attttctgct
     3181 cactatagag aaaggggaag cagaacatct aagaaagagg gacaagctgt ggggggtctg
     3241 tgtctccatc ctggctctct tgcctcgagt cctcaggttg atgctgcaga gcctgcgggt
     3301 gaacagagtt gggcctgagg agctgcctgt tgtgggccag ctgcttcgac tgctgcttca
     3361 gcatgcaccc ctcaggactc atatgttgac caatgcgatc ttggtgcagc agatcatcaa
     3421 gaatatcacg acattgaaga gtggaagtgt tcaggaacag tggctcacag acttacatta
     3481 ctgcttcaac gtgtatatca ctgggcatcc ccaagggccc agtgcactgg ctacagtgta
     3541 tctcgagtct agagactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3601 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     3661 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3721 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3781 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3841 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3901 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3961 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4021 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4081 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4141 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4201 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     4261 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4321 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4381 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4441 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4501 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     4561 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4621 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4681 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     4741 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4801 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4861 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     4921 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4981 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5041 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5161 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5221 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5281 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5341 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5401 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5461 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5521 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5581 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5641 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5701 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5761 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5821 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5881 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5941 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6001 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6061 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6121 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6181 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6241 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6301 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6481 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6541 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6601 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6661 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6721 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6901 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7081 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7201 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7261 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7321 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7381 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7441 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7501 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7561 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7621 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7681 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7741 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7801 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7861 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7921 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7981 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8041 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8101 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8161 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8221 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8281 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8341 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8401 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8461 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8521 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8581 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8641 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8701 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8761 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8821 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
