LOCUS       pEnCMV-GK(human)-HA     7636 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036703
VERSION     V036703
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             706..2295
                     /label="GK(NM_203391)"
                     /note="GK(NM_203391)"
                     /gene="GK"
     CDS             2302..2328
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2332..2358
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2362..2388
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2412..3000
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3066..3114
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3316..3744
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3758..4087
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4154..4948
                     /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    5124..5245
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5294..5310)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5318..5334
                     /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(5342..5372)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5387..5408
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5696..6284)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6455..7315)
                     /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(7316..7420)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc agcctcaaag
      721 aaggcagttt tggggccatt ggtgggggcg gtggaccagg gcaccagttc gacgcgcttt
      781 ttggttttca attcaaaaac agctgaacta cttagtcatc atcaagtaga aataaaacaa
      841 gagttcccaa gagaaggatg ggtggaacag gaccctaagg aaattctaca ttctgtctat
      901 gagtgtatag agaaaacatg tgagaaactt ggacagctca atattgatat ttccaacata
      961 aaagctattg gtgtcagcaa ccagagggaa accactgtag tctgggacaa gataactgga
     1021 gagcctctct acaatgctgt ggtgtggctt gatctaagaa cccagtctac cgttgagagt
     1081 cttagtaaaa gaattccagg aaataataac tttgtcaagt ccaagacagg ccttccactt
     1141 agcacttact tcagtgcagt gaaacttcgt tggctccttg acaatgtgag aaaagttcaa
     1201 aaggccgttg aagaaaaacg agctcttttt gggactattg attcatggct tatttggagt
     1261 ttgacaggag gagtcaatgg aggtgtccac tgtacagatg taacaaatgc aagtaggact
     1321 atgcttttca acattcattc tttggaatgg gataaacaac tctgcgaatt ttttggaatt
     1381 ccaatggaaa ttcttccaaa tgtccggagt tcttctgaga tctatggcct aatgaaaatc
     1441 tctcatagcg tgaaagctgg ggccttggaa ggtgtgccaa tatctgggtg tttaggggac
     1501 cagtctgctg cattggtggg acaaatgtgc ttccagattg gacaagccaa aaatacgtat
     1561 ggaacaggat gtttcttact atgtaataca ggccataagt gtgtattttc tgatcatggc
     1621 cttctcacca cagtggctta caaacttggc agagacaaac cagtatatta tgctttggaa
     1681 ggttctgtag ctatagctgg tgctgttatt cgctggctaa gagacaatct tggaattata
     1741 aagacctcag aagaaattga aaaacttgct aaagaagtag gtacttctta tggctgctac
     1801 ttcgtcccag cattttcggg gttatatgca ccttattggg agcccagcgc aagagggata
     1861 atctgtggac tcactcagtt caccaataaa tgccatattg cttttgctgc attagaagct
     1921 gtttgtttcc aaactcgaga gattttggat gccatgaatc gagactgtgg aattccactc
     1981 agtcatttgc aggtagatgg aggaatgacc agcaacaaaa ttcttatgca gctacaagca
     2041 gacattctgt atataccagt agtgaagccc tcaatgcccg aaaccactgc actgggtgcg
     2101 gctatggcgg caggggctgc agaaggagtc ggcgtatgga gtctcgaacc cgaggatttg
     2161 tctgccgtca cgatggagcg gtttgaacct cagattaatg cggaggaaag tgaaattcgt
     2221 tattctacat ggaagaaagc tgtgatgaag tcaatgggtt gggttacaac tcaatctcca
     2281 gaaagtggta ttccactcga gtacccatac gacgtacctg attatgcaac ttatccttat
     2341 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata gtgagggccc
     2401 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2461 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2521 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2581 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2641 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2701 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2761 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     2821 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2881 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2941 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3001 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     3061 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     3121 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     3181 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     3241 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     3301 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3361 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3421 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     3481 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3541 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3601 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3661 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     3721 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3781 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3841 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3901 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3961 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4021 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4081 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4141 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4201 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4261 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4321 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4381 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4441 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4501 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4561 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4621 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4681 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4741 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4801 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4861 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4921 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     4981 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5041 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5101 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5161 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5221 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5281 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5341 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5401 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5461 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5521 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5581 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5641 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5881 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6121 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7441 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     7501 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7561 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7621 gaagaatctg cttagg
//
