LOCUS       V037222                 9631 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037222
VERSION     V037222
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1030..5151
                     /label="MAP3K5(NM_005923)"
                     /note="MAP3K5(NM_005923)"
                     /gene="MAP3K5"
     polyA_signal    5231..5455
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5501..5929
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5943..6272
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6339..7133
                     /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    7307..7428
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7477..7493)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7501..7517
                     /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(7525..7555)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7570..7591
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7879..8464)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8635..9495)
                     /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(9496..9600)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
     1021 gaaaagctta tgagcacgga ggcggacgag ggcatcactt tctctgtgcc acccttcgcc
     1081 ccctcgggct tctgcaccat ccccgagggc ggcatctgca ggaggggagg agcggcggcg
     1141 gtgggcgagg gcgaggagca ccagctgcca ccgccgccgc cgggcagctt ctggaacgtg
     1201 gagagcgccg ctgcccctgg catcggttgt ccggcggcca cctcctcgag cagtgccacc
     1261 cgaggccggg gcagctctgt tggcgggggc agccgacgga ccacggtggc atatgtgatc
     1321 aacgaagcga gccaagggca actggtggtg gccgagagcg aggccctgca gagcttgcgg
     1381 gaggcgtgcg agacagtggg cgccaccctg gaaaccctgc attttgggaa actcgacttt
     1441 ggagaaacca ccgtgctgga ccgcttttac aatgcagata ttgcggtggt ggagatgagc
     1501 gatgccttcc ggcagccgtc cttgttttac caccttgggg tgagagaaag tttcagcatg
     1561 gccaacaaca tcatcctcta ctgtgatact aactcggact ctctgcagtc actgaaggaa
     1621 ataatttgcc agaagaatac tatgtgcact gggaactaca cctttgttcc ttacatgata
     1681 actccacata acaaagtcta ctgctgtgac agcagcttca tgaaggggtt gacagagctc
     1741 atgcaaccga acttcgagct gcttcttgga cccatctgct tacctcttgt ggatcgtttt
     1801 attcaacttt tgaaggtggc acaagcaagt tctagccagt acttccggga atctatactc
     1861 aatgacatca ggaaagctcg taatttatac actggtaaag aattggcagc tgagttggca
     1921 agaattcggc agcgagtaga taatatcgaa gtcttgacag cagatattgt cataaatctg
     1981 ttactttcct acagagatat ccaggactat gattctattg tgaagctggt agagacttta
     2041 gaaaaactgc caacctttga tttggcctcc catcaccatg tgaagtttca ttatgcattt
     2101 gcactgaata ggagaaatct ccctggtgac agagcaaaag ctcttgatat tatgattccc
     2161 atggtgcaaa gcgaaggaca agttgcttca gatatgtatt gcctagttgg tcgaatctac
     2221 aaagatatgt ttttggactc taatttcacg gacactgaaa gcagagacca tggagcttct
     2281 tggttcaaaa aggcatttga atctgagcca acactacagt caggaattaa ttatgcggtc
     2341 ctcctcctgg cagctggaca ccagtttgaa tcttcctttg agctccggaa agttggggtg
     2401 aagctaagta gtcttcttgg taaaaaggga aacttggaaa aactccagag ctactgggaa
     2461 gttggatttt ttctgggggc cagcgtccta gccaatgacc acatgagagt cattcaagca
     2521 tctgaaaagc tttttaaact gaagacacca gcatggtacc tcaagtctat tgtagagaca
     2581 attttaatat ataagcattt tgtgaaactg accacagaac agcctgtggc caagcaagaa
     2641 cttgtggact tttggatgga tttcctggtc gaggccacaa agacagatgt tactgtggtt
     2701 aggtttccag tattaatatt agaaccaacc aaaatctatc aaccttctta tttgtctatc
     2761 aacaatgaag ttgaggaaaa gacaatctct atttggcacg tgcttcctga tgacaagaaa
     2821 ggtatacatg agtggaattt tagtgcctct tctgtcaggg gagtgagtat ttctaaattt
     2881 gaagaaagat gctgctttct ttatgtgctt cacaattctg atgatttcca aatctatttc
     2941 tgtacagaac ttcattgtaa aaagtttttt gagatggtga acaccattac cgaagagaag
     3001 gggagaagca cagaggaagg agactgtgaa agtgacttgc tggagtatga ctatgaatat
     3061 gatgaaaatg gtgacagagt cgttttagga aaaggcactt atgggatagt ctacgcaggt
     3121 cgggacttga gcaaccaagt cagaattgct attaaggaaa tcccagagag agacagcaga
     3181 tactctcagc ccctgcatga agaaatagca ttgcataaac acctgaagca caaaaatatt
     3241 gtccagtatc tgggctcttt cagtgagaat ggtttcatta aaatcttcat ggagcaggtc
     3301 cctggaggaa gtctttctgc tctccttcgt tccaaatggg gtccattaaa agacaatgag
     3361 caaacaattg gcttttatac aaagcaaata ctggaaggat taaaatatct ccatgacaat
     3421 cagatagttc accgggacat aaagggtgac aatgtgttga ttaataccta cagtggtgtt
     3481 ctcaagatct ctgacttcgg aacatcaaag aggcttgctg gcataaaccc ctgtactgaa
     3541 acttttactg gtaccctcca gtatatggca ccagaaataa tagataaagg accaagaggc
     3601 tacggaaaag cagcagacat ctggtctctg ggctgtacaa tcattgaaat ggccacagga
     3661 aaacccccat tttatgaact gggagaacca caagcagcta tgttcaaggt gggaatgttt
     3721 aaagtccacc ctgagatccc agagtccatg tctgcagagg ccaaggcatt catactgaaa
     3781 tgttttgaac cagatcctga caagagagcc tgtgctaacg acttgcttgt tgatgagttt
     3841 ttaaaagttt caagcaaaaa gaaaaagaca caacctaagc tttcagctct ttcagctgga
     3901 tcaaatgaat atctcaggag tatatccttg ccggtacctg tgctggtgga ggacaccagc
     3961 agcagcagtg agtacggctc agtttcaccc gacacggagt tgaaagtgga ccccttctct
     4021 ttcaaaacaa gagccaagtc ctgcggagaa agagatgtca agggaattcg gacactcttt
     4081 ttgggcattc cagatgagaa ttttgaagat cacagtgctc ctccttcccc tgaagaaaaa
     4141 gattctggat tcttcatgct gaggaaggac agtgagaggc gagctaccct tcacaggatc
     4201 ctgacggaag accaagacaa aattgtgaga aacctaatgg aatctttagc tcagggggct
     4261 gaagaaccga aactaaaatg ggaacacatc acaaccctca ttgcaagcct cagagaattt
     4321 gtgagatcca ctgaccgaaa aatcatagcc accacactgt caaagctgaa actggagctg
     4381 gacttcgaca gccatggcat tagccaagtc caggtggtac tctttggttt tcaagatgct
     4441 gtcaataaag ttcttcggaa tcataacatc aagccgcact ggatgtttgc cttagacagt
     4501 atcattcgga aggcggtaca gacagccatt accatcctgg ttccagaact aaggccacat
     4561 ttcagccttg catctgagag tgatactgct gatcaagaag acttggatgt agaagatgac
     4621 catgaggaac agccttcaaa tcaaactgtc cgaagacctc aggctgtcat tgaagatgct
     4681 gtggctacct caggcgtgag cacgctcagt tctactgtgt ctcatgattc ccagagtgct
     4741 caccggtcac tgaatgtaca gcttggaagg atgaaaatag aaaccaatag attactggaa
     4801 gaattggttc ggaaagagaa agaattacaa gcactccttc atcgagctat tgaagaaaaa
     4861 gaccaagaaa ttaaacacct gaagcttaag tcccaaccca tagaaattcc tgaattgcct
     4921 gtatttcatc taaattcttc tggcacaaat actgaagatt ctgaacttac cgactggctg
     4981 agagtgaatg gagctgatga agacactata agccggtttt tggctgaaga ttatacacta
     5041 ttggatgttc tctactatgt tacacgtgat gacttaaaat gcttgagact aaggggaggg
     5101 atgctgtgca cactgtggaa ggctatcatt gactttcgaa acaaacagac ttgagaattc
     5161 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     5221 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     5281 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     5341 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     5401 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     5461 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     5521 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5581 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5641 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5701 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5761 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5821 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5881 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6181 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6241 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     6301 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6361 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6421 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6481 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6541 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6601 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6661 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6721 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6781 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6841 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6901 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6961 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     7021 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     7081 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     7141 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7201 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7261 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7321 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7381 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7441 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7501 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7561 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7621 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7681 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8401 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8761 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9601 cacatttccc cgaaaagtgc cacctgacgt c
//
