LOCUS       V045977                 8433 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045977
VERSION     V045977
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..3113
                     /label="Stat5b(NM_011489)"
                     /note="Stat5b(NM_011489)"
                     /gene="Stat5b"
     CDS             3120..3185
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3209..3797
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3863..3911
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4113..4541
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4555..4884
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4951..5745
                     /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    5921..6042
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6091..6107)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6115..6131
                     /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(6139..6169)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6184..6205
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6493..7081)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7252..8112)
                     /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(8113..8217)
                     /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 tatgtggata caggctcagc
      781 agctccaggg cgatgccctt caccagatgc aggccttgta cggccagcat ttccccatcg
      841 aggtgcgaca ttatttatca cagtggatcg aaagccaagc ctgggactca atagatcttg
      901 ataatccaca ggagaacatt aaggccaccc agctcctgga gggcctggtg caggagctgc
      961 agaagaaggc ggagcaccag gtgggggaag atgggttttt gctgaagatc aagctggggc
     1021 actatgccac acagctccag agcacgtacg accgctgccc catggagctg gttcgctgta
     1081 tccggcacat tctgtacaac gaacagaggc tggttcgcga agccaacaac ggcagctctc
     1141 cagctggaag tcttgctgac gccatgtccc agaagcacct tcagatcaac caaacgtttg
     1201 aggagctgcg cctgatcaca caggacacgg agaacgagct gaagaagctg cagcagaccc
     1261 aagagtactt catcatccag taccaggaga gcctgcggat ccaagctcag tttgcccagc
     1321 tgggacagct gaacccccag gagcgcatga gcagggagac ggccctccag cagaagcaag
     1381 tgtccctgga gacctggctg cagcgagagg cacagacact gcagcagtac cgagtggagc
     1441 tggctgagaa gcaccagaag accctgcagc tgctgcggaa gcagcagacc atcatcctgg
     1501 acgacgagct gatccagtgg aagcggagac agcagctggc cgggaacggg ggtccccccg
     1561 agggcagcct ggacgtgctg cagtcctggt gtgagaagct ggccgagatc atctggcaga
     1621 accggcagca gatccgcagg gctgagcact tgtgccagca gctgcccatc ccaggccccg
     1681 tggaggagat gctggctgag gtcaacgcca ccatcacgga catcatctca gccctggtca
     1741 ccagcacgtt catcatcgag aagcagcctc ctcaggtcct gaagacccag accaagtttg
     1801 cagccaccgt gcgcctgctg gtggggggga agctgaatgt gcacatgaac cccccgcagg
     1861 tgaaggcgac catcatcagc gagcagcagg ccaagtccct gctcaagaat gagaacaccc
     1921 gcaatgatta cagcggcgag atcctgaaca actgttgcgt catggagtac caccaggcca
     1981 ctggcacact cagcgcccac ttcagaaaca tgtccctgaa acgaatcaag aggtctgacc
     2041 gccgtggggc agagtcagta acggaagaga agttcacgat cctgtttgac tcacagttca
     2101 gcgtcggtgg aaacgagctg gtctttcaag tcaagacctt gtcgctcccg gtggtggtga
     2161 ttgttcacgg cagccaggac aacaatgcca cagccactgt cctctgggac aacgcctttg
     2221 cagagcctgg cagggtgcca tttgccgtgc ctgacaaggt gctgtggccg cagctgtgtg
     2281 aagcgctcaa catgaaattc aaggctgaag tacagagcaa ccggggcttg accaaggaga
     2341 acctcgtgtt cctggcacag aaactgttca acatcagcag caaccacctc gaggactaca
     2401 acagcatgtc cgtgtcctgg tcccagttca accgggagaa tttgccagga cggaattaca
     2461 ctttctggca gtggtttgat ggcgtgatgg aagtattgaa aaaacatctc aagcctcact
     2521 ggaatgatgg ggctatcctg ggtttcgtga acaagcaaca ggcccacgac ctgctcatca
     2581 acaagccaga cgggaccttc ctgctgcgct tcagcgactc ggaaatcggg ggcatcacca
     2641 ttgcttggaa gtttgactct caggagagaa tgttttggaa tctgatgcct tttaccacta
     2701 gagacttctc tatccggtcc ctcgctgacc gcctggggga cctgaattac ctcatatatg
     2761 tgtttcctga tcggccaaag gatgaagtat attctaagta ctacacaccg gtcccctgtg
     2821 agcccgcaac tgcgaaagca gctgacggat acgtgaagcc acagatcaag caggtggtcc
     2881 ccgagtttgc aaatgcatcc acagatgctg ggagtggcgc cacctacatg gatcaggctc
     2941 cttccccagt cgtgtgccct caggctcact acaacatgta cccacccaac ccggactccg
     3001 tccttgatac cgatggggac ttcgatctgg aagacacgat ggacgtggcg cggcgcgtgg
     3061 aagagctctt aggccggccc atggacagtc agtggatccc tcacgcacag tcaggatccg
     3121 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     3181 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3241 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3301 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3361 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3421 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3481 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3541 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3601 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3661 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3721 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3781 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     3841 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3901 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3961 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4021 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     4081 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4141 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4201 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4261 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4321 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4381 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4441 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4501 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4561 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4621 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4681 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4741 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4801 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4861 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4921 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4981 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5041 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5101 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5161 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5221 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5281 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5341 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5401 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5461 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5521 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5581 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5641 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5701 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5761 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5821 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5881 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6001 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6061 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6121 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6181 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6241 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6301 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6361 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6421 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6481 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6541 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6601 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6661 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6721 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6781 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6841 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6901 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6961 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7021 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7081 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7141 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7201 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7261 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7321 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7381 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7441 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7501 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7561 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7621 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7681 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7741 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7801 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7861 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7921 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7981 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8041 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8101 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8161 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8221 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8281 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     8341 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     8401 ttgaccgaca attgcatgaa gaatctgctt agg
//
