LOCUS       V021391                 9497 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021391
VERSION     V021391
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"
     CDS             683..4144
                     /label="JAK1(NM_001320923)"
                     /note="JAK1(NM_001320923)"
                     /gene="JAK1"
     CDS             4184..4249
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4273..4861
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4927..4975
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5177..5605
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5619..5948
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6015..6809
                     /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    6985..7106
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7155..7171)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7179..7195
                     /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(7203..7233)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7248..7269
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7557..8145)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8316..9176)
                     /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(9177..9281)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgcagta tctaaatata aaagaggact gcaatgccat
      721 ggctttctgt gctaaaatga ggagctccaa gaagactgag gtgaacctgg aggcccctga
      781 gccaggggtg gaagtgatct tctatctgtc ggacagggag cccctccggc tgggcagtgg
      841 agagtacaca gcagaggaac tgtgcatcag ggctgcacag gcatgccgta tctctcctct
      901 ttgtcacaac ctctttgccc tgtatgacga gaacaccaag ctctggtatg ctccaaatcg
      961 caccatcacc gttgatgaca agatgtccct ccggctccac taccggatga ggttctattt
     1021 caccaattgg catggaacca acgacaatga gcagtcagtg tggcgtcatt ctccaaagaa
     1081 gcagaaaaat ggctacgaga aaaaaaagat tccagatgca acccctctcc ttgatgccag
     1141 ctcactggag tatctgtttg ctcagggaca gtatgatttg gtgaaatgcc tggctcctat
     1201 tcgagacccc aagaccgagc aggatggaca tgatattgag aacgagtgtc tagggatggc
     1261 tgtcctggcc atctcacact atgccatgat gaagaagatg cagttgccag aactgcccaa
     1321 ggacatcagc tacaagcgat atattccaga aacattgaat aagtccatca gacagaggaa
     1381 ccttctcacc aggatgcgga taaataatgt tttcaaggat ttcctaaagg aatttaacaa
     1441 caagaccatt tgtgacagca gcgtgtccac gcatgacctg aaggtgaaat acttggctac
     1501 cttggaaact ttgacaaaac attacggtgc tgaaatattt gagacttcca tgttactgat
     1561 ttcatcagaa aatgagatga attggtttca ttcgaatgac ggtggaaacg ttctctacta
     1621 cgaagtgatg gtgactggga atcttggaat ccagtggagg cataaaccaa atgttgtttc
     1681 tgttgaaaag gaaaaaaata aactgaagcg gaaaaaactg gaaaataaac acaagaagga
     1741 tgaggagaaa aacaagatcc gggaagagtg gaacaatttt tcttacttcc ctgaaatcac
     1801 tcacattgta ataaaggagt ctgtggtcag cattaacaag caggacaaca agaaaatgga
     1861 actgaagctc tcttcccacg aggaggcctt gtcctttgtg tccctggtag atggctactt
     1921 ccggctcaca gcagatgccc atcattacct ctgcaccgac gtggcccccc cgttgatcgt
     1981 ccacaacata cagaatggct gtcatggtcc aatctgtaca gaatacgcca tcaataaatt
     2041 gcggcaagaa ggaagcgagg aggggatgta cgtgctgagg tggagctgca ccgactttga
     2101 caacatcctc atgaccgtca cctgctttga gaagtctgag caggtgcagg gtgcccagaa
     2161 gcagttcaag aactttcaga tcgaggtgca gaagggccgc tacagtctgc acggttcgga
     2221 ccgcagcttc cccagcttgg gagacctcat gagccacctc aagaagcaga tcctgcgcac
     2281 ggataacatc agcttcatgc taaaacgctg ctgccagccc aagccccgag aaatctccaa
     2341 cctgctggtg gctactaaga aagcccagga gtggcagccc gtctacccca tgagccagct
     2401 gagtttcgat cggatcctca agaaggatct ggtgcagggc gagcaccttg ggagaggcac
     2461 gagaacacac atctattctg ggaccctgat ggattacaag gatgacgaag gaacttctga
     2521 agagaagaag ataaaagtga tcctcaaagt cttagacccc agccacaggg atatttccct
     2581 ggccttcttc gaggcagcca gcatgatgag acaggtctcc cacaaacaca tcgtgtacct
     2641 ctatggcgtc tgtgtccgcg acgtggagaa tatcatggtg gaagagtttg tggaaggggg
     2701 tcctctggat ctcttcatgc accggaaaag cgatgtcctt accacaccat ggaaattcaa
     2761 agttgccaaa cagctggcca gtgccctgag ctacttggag gataaagacc tggtccatgg
     2821 aaatgtgtgt actaaaaacc tcctcctggc ccgtgagggc atcgacagtg agtgtggccc
     2881 attcatcaag ctcagtgacc ccggcatccc cattacggtg ctgtctaggc aagaatgcat
     2941 tgaacgaatc ccatggattg ctcctgagtg tgttgaggac tccaagaacc tgagtgtggc
     3001 tgctgacaag tggagctttg gaaccacgct ctgggaaatc tgctacaatg gcgagatccc
     3061 cttgaaagac aagacgctga ttgagaaaga gagattctat gaaagccggt gcaggccagt
     3121 gacaccatca tgtaaggagc tggctgacct catgacccgc tgcatgaact atgaccccaa
     3181 tcagaggcct ttcttccgag ccatcatgag agacattaat aagcttgaag agcagaatcc
     3241 agatattgtt tcagaaaaaa aaccagcaac tgaagtggac cccacacatt ttgaaaagcg
     3301 cttcctaaag aggatccgtg acttgggaga gggccacttt gggaaggttg agctctgcag
     3361 gtatgacccc gaaggggaca atacagggga gcaggtggct gttaaatctc tgaagcctga
     3421 gagtggaggt aaccacatag ctgatctgaa aaaggaaatc gagatcttaa ggaacctcta
     3481 tcatgagaac attgtgaagt acaaaggaat ctgcacagaa gacggaggaa atggtattaa
     3541 gctcatcatg gaatttctgc cttcgggaag ccttaaggaa tatcttccaa agaataagaa
     3601 caaaataaac ctcaaacagc agctaaaata tgccgttcag atttgtaagg ggatggacta
     3661 tttgggttct cggcaatacg ttcaccggga cttggcagca agaaatgtcc ttgttgagag
     3721 tgaacaccaa gtgaaaattg gagacttcgg tttaaccaaa gcaattgaaa ccgataagga
     3781 gtattacacc gtcaaggatg accgggacag ccctgtgttt tggtatgctc cagaatgttt
     3841 aatgcaatct aaattttata ttgcctctga cgtctggtct tttggagtca ctctgcatga
     3901 gctgctgact tactgtgatt cagattctag tcccatggct ttgttcctga aaatgatagg
     3961 cccaacccat ggccagatga cagtcacaag acttgtgaat acgttaaaag aaggaaaacg
     4021 cctgccgtgc ccacctaact gtccagatga ggtttatcaa cttatgagga aatgctggga
     4081 attccaacca tccaatcgga caagctttca gaaccttatt gaaggatttg aagcactttt
     4141 aaaagaattc tgcagatatc cagcacagtg gcggccgctc gaggactaca aagaccatga
     4201 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     4261 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4321 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4381 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4441 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4501 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4561 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4621 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4681 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4741 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4801 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4861 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4921 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     4981 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5041 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5101 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     5161 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5221 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5281 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     5341 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5401 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5461 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5521 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     5581 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5641 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5701 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5761 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5821 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5881 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5941 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     6001 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6061 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6121 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6181 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6241 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6301 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6361 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6421 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6481 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6541 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6601 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6661 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6721 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6781 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     6841 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6901 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6961 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     7021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7081 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     7141 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7201 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7261 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7741 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7981 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9301 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     9361 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     9421 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     9481 tgaagaatct gcttagg
//
