LOCUS       V042515                 7564 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042515
VERSION     V042515
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             709..726
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             727..2223
                     /label="CAMK2D(NM_001221)"
                     /note="CAMK2D(NM_001221)"
                     /gene="CAMK2D"
     CDS             2230..2256
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2260..2286
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2290..2316
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2340..2928
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2994..3042
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3244..3672
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3686..4015
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4082..4876
                     /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    5052..5173
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5222..5238)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5246..5262
                     /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(5270..5300)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5315..5336
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5624..6212)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6383..7243)
                     /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(7244..7348)
                     /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 ccaccatgca tcatcatcat
      721 catcacatgg cttcgaccac aacctgcacc aggttcacgg acgagtatca gcttttcgag
      781 gagcttggaa agggggcatt ctcagtggtg agaagatgta tgaaaattcc tactggacaa
      841 gaatatgctg ccaaaattat caacaccaaa aagctttctg ctagggatca tcagaaacta
      901 gaaagagaag ctagaatctg ccgtcttttg aagcacccta atattgtgcg acttcatgat
      961 agcatatcag aagagggctt tcactacttg gtgtttgatt tagttactgg aggtgaactg
     1021 tttgaagaca tagtggcaag agaatactac agtgaagctg atgccagtca ttgtatacag
     1081 cagattctag aaagtgttaa tcattgtcac ctaaatggca tagttcacag ggacctgaag
     1141 cctgagaatt tgcttttagc tagcaaatcc aagggagcag ctgtgaaatt ggcagacttt
     1201 ggcttagcca tagaagttca aggggaccag caggcgtggt ttggttttgc tggcacacct
     1261 ggatatcttt ctccagaagt tttacgtaaa gatccttatg gaaagccagt ggatatgtgg
     1321 gcatgtggtg tcattctcta tattctactt gtggggtatc cacccttctg ggatgaagac
     1381 caacacagac tctatcagca gatcaaggct ggagcttatg attttccatc accagaatgg
     1441 gacacggtga ctcctgaagc caaagacctc atcaataaaa tgcttactat caaccctgcc
     1501 aaacgcatca cagcctcaga ggcactgaag cacccatgga tctgtcaacg ttctactgtt
     1561 gcttccatga tgcacagaca ggagactgta gactgcttga agaaatttaa tgctagaaga
     1621 aaactaaagg gtgccatctt gacaactatg ctggctacaa ggaatttctc agcagccaag
     1681 agtttgttga agaaaccaga tggagtaaag gagtcaactg agagttcaaa tacaacaatt
     1741 gaggatgaag atgtgaaagc acgaaagcaa gagattatca aagtcactga acaactgatc
     1801 gaagctatca acaatgggga ctttgaagcc tacacaaaaa tctgtgaccc aggccttact
     1861 gcttttgaac ctgaagcttt gggtaattta gtggaaggga tggattttca ccgattctac
     1921 tttgaaaatg ctttgtccaa aagcaataaa ccaatccaca ctattattct aaaccctcat
     1981 gtacatctgg taggggatga tgccgcctgc atagcatata ttaggctcac acagtacatg
     2041 gatggcagtg gaatgccaaa gacaatgcag tcagaagaga ctcgtgtgtg gcaccgccgg
     2101 gatggaaagt ggcagaatgt tcattttcat cgctcggggt caccaacagt acccatcaag
     2161 ccaccctgta ttccaaatgg gaaagaaaac ttctcaggag gcacctcttt gtggcaaaac
     2221 atcctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat
     2281 tatgctagct atccttatga tgtcccagac tacgcatagt gagggcccga tatctcgaca
     2341 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2401 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2461 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2521 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2581 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2641 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2701 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     2761 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2821 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2881 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     2941 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3001 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3061 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3121 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3181 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     3241 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3301 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3361 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     3421 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3481 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3541 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3601 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3661 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3781 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3841 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3901 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3961 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4021 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4081 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4141 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4201 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4261 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4321 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4381 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4441 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4501 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4561 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4621 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4681 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4741 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4801 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4861 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     4921 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4981 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5041 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5161 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5221 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5281 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5341 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5401 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5461 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5521 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5581 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5641 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5701 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5761 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     5821 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5881 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5941 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6001 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6061 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6121 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6181 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6241 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6301 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6361 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6421 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6481 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6541 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6601 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6661 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6721 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6781 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6841 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6901 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6961 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7021 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7081 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7141 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7201 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7261 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7321 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7381 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     7441 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7501 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7561 tagg
//
