LOCUS       V043166                10269 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043166
VERSION     V043166
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..4217
                     /label="JAK1(NM_002227)"
                     /note="JAK1(NM_002227)"
                     /gene="JAK1"
     CDS             4233..4949
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced CFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG
                     IKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="ECFP"
     CDS             4956..5021
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5045..5633
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5699..5747
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5949..6377
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6391..6720
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6787..7581
                     /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    7757..7878
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7927..7943)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7951..7967
                     /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(7975..8005)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8020..8041
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8329..8917)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9088..9948)
                     /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(9949..10053)
                     /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 ccaccatgca gtatctaaat ataaaagagg
      781 actgcaatgc catggctttc tgtgctaaaa tgaggagctc caagaagact gaggtgaacc
      841 tggaggcccc tgagccaggg gtggaagtga tcttctatct gtcggacagg gagcccctcc
      901 ggctgggcag tggagagtac acagcagagg aactgtgcat cagggctgca caggcatgcc
      961 gtatctctcc tctttgtcac aacctctttg ccctgtatga cgagaacacc aagctctggt
     1021 atgctccaaa tcgcaccatc accgttgatg acaagatgtc cctccggctc cactaccgga
     1081 tgaggttcta tttcaccaat tggcatggaa ccaacgacaa tgagcagtca gtgtggcgtc
     1141 attctccaaa gaagcagaaa aatggctacg agaaaaaaaa gattccagat gcaacccctc
     1201 tccttgatgc cagctcactg gagtatctgt ttgctcaggg acagtatgat ttggtgaaat
     1261 gcctggctcc tattcgagac cccaagaccg agcaggatgg acatgatatt gagaacgagt
     1321 gtctagggat ggctgtcctg gccatctcac actatgccat gatgaagaag atgcagttgc
     1381 cagaactgcc caaggacatc agctacaagc gatatattcc agaaacattg aataagtcca
     1441 tcagacagag gaaccttctc accaggatgc ggataaataa tgttttcaag gatttcctaa
     1501 aggaatttaa caacaagacc atttgtgaca gcagcgtgtc cacgcatgac ctgaaggtga
     1561 aatacttggc taccttggaa actttgacaa aacattacgg tgctgaaata tttgagactt
     1621 ccatgttact gatttcatca gaaaatgaga tgaattggtt tcattcgaat gacggtggaa
     1681 acgttctcta ctacgaagtg atggtgactg ggaatcttgg aatccagtgg aggcataaac
     1741 caaatgttgt ttctgttgaa aaggaaaaaa ataaactgaa gcggaaaaaa ctggaaaata
     1801 aacacaagaa ggatgaggag aaaaacaaga tccgggaaga gtggaacaat ttttcttact
     1861 tccctgaaat cactcacatt gtaataaagg agtctgtggt cagcattaac aagcaggaca
     1921 acaagaaaat ggaactgaag ctctcttccc acgaggaggc cttgtccttt gtgtccctgg
     1981 tagatggcta cttccggctc acagcagatg cccatcatta cctctgcacc gacgtggccc
     2041 ccccgttgat cgtccacaac atacagaatg gctgtcatgg tccaatctgt acagaatacg
     2101 ccatcaataa attgcggcaa gaaggaagcg aggaggggat gtacgtgctg aggtggagct
     2161 gcaccgactt tgacaacatc ctcatgaccg tcacctgctt tgagaagtct gagcaggtgc
     2221 agggtgccca gaagcagttc aagaactttc agatcgaggt gcagaagggc cgctacagtc
     2281 tgcacggttc ggaccgcagc ttccccagct tgggagacct catgagccac ctcaagaagc
     2341 agatcctgcg cacggataac atcagcttca tgctaaaacg ctgctgccag cccaagcccc
     2401 gagaaatctc caacctgctg gtggctacta agaaagccca ggagtggcag cccgtctacc
     2461 ccatgagcca gctgagtttc gatcggatcc tcaagaagga tctggtgcag ggcgagcacc
     2521 ttgggagagg cacgagaaca cacatctatt ctgggaccct gatggattac aaggatgacg
     2581 aaggaacttc tgaagagaag aagataaaag tgatcctcaa agtcttagac cccagccaca
     2641 gggatatttc cctggccttc ttcgaggcag ccagcatgat gagacaggtc tcccacaaac
     2701 acatcgtgta cctctatggc gtctgtgtcc gcgacgtgga gaatatcatg gtggaagagt
     2761 ttgtggaagg gggtcctctg gatctcttca tgcaccggaa aagcgatgtc cttaccacac
     2821 catggaaatt caaagttgcc aaacagctgg ccagtgccct gagctacttg gaggataaag
     2881 acctggtcca tggaaatgtg tgtactaaaa acctcctcct ggcccgtgag ggcatcgaca
     2941 gtgagtgtgg cccattcatc aagctcagtg accccggcat ccccattacg gtgctgtcta
     3001 ggcaagaatg cattgaacga atcccatgga ttgctcctga gtgtgttgag gactccaaga
     3061 acctgagtgt ggctgctgac aagtggagct ttggaaccac gctctgggaa atctgctaca
     3121 atggcgagat ccccttgaaa gacaagacgc tgattgagaa agagagattc tatgaaagcc
     3181 ggtgcaggcc agtgacacca tcatgtaagg agctggctga cctcatgacc cgctgcatga
     3241 actatgaccc caatcagagg cctttcttcc gagccatcat gagagacatt aataagcttg
     3301 aagagcagaa tccagatatt gtttcagaaa aaaaaccagc aactgaagtg gaccccacac
     3361 attttgaaaa gcgcttccta aagaggatcc gtgacttggg agagggccac tttgggaagg
     3421 ttgagctctg caggtatgac cccgaagggg acaatacagg ggagcaggtg gctgttaaat
     3481 ctctgaagcc tgagagtgga ggtaaccaca tagctgatct gaaaaaggaa atcgagatct
     3541 taaggaacct ctatcatgag aacattgtga agtacaaagg aatctgcaca gaagacggag
     3601 gaaatggtat taagctcatc atggaatttc tgccttcggg aagccttaag gaatatcttc
     3661 caaagaataa gaacaaaata aacctcaaac agcagctaaa atatgccgtt cagatttgta
     3721 aggggatgga ctatttgggt tctcggcaat acgttcaccg ggacttggca gcaagaaatg
     3781 tccttgttga gagtgaacac caagtgaaaa ttggagactt cggtttaacc aaagcaattg
     3841 aaaccgataa ggagtattac accgtcaagg atgaccggga cagccctgtg ttttggtatg
     3901 ctccagaatg tttaatgcaa tctaaatttt atattgcctc tgacgtctgg tcttttggag
     3961 tcactctgca tgagctgctg acttactgtg attcagattc tagtcccatg gctttgttcc
     4021 tgaaaatgat aggcccaacc catggccaga tgacagtcac aagacttgtg aatacgttaa
     4081 aagaaggaaa acgcctgccg tgcccaccta actgtccaga tgaggtttat caacttatga
     4141 ggaaatgctg ggaattccaa ccatccaatc ggacaagctt tcagaacctt attgaaggat
     4201 ttgaagcact tttaaaagga ggaggtggat ccatggtgag caagggcgag gagctgttca
     4261 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4321 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4381 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc
     4441 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4501 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     4561 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     4621 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca
     4681 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     4741 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4801 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     4861 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4921 tcactctcgg catggacgag ctgtacaagg gatccgacta caaagaccat gacggtgatt
     4981 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     5041 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5101 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5161 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5221 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5281 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5341 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5401 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5461 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5521 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5581 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     5641 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5701 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     5761 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5821 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5881 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     5941 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6001 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6061 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     6121 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6181 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6241 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6301 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     6361 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6421 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6541 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6601 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     6661 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6721 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     6781 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6841 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6901 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6961 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     7021 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     7081 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7141 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7201 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7261 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7321 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7381 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7441 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7501 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7561 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     7621 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7681 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7741 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7801 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7861 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7921 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7981 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     8041 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8101 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     8161 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8221 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8281 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8521 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8701 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8761 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8821 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     9001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9061 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     9121 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9181 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9241 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9301 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9361 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9421 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9481 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9541 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9601 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9661 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9721 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9781 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9841 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9901 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9961 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    10021 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    10081 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
    10141 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    10201 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    10261 ctgcttagg
//
