LOCUS       V031795                 6976 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031795
VERSION     V031795
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..1632
                     /label="PPP2CA(NM_002715)"
                     /note="PPP2CA(NM_002715)"
                     /gene="PPP2CA"
     CDS             1639..1728
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1752..2340
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2406..2454
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2656..3084
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3098..3427
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3494..4288
                     /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    4464..4585
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4634..4650)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4658..4674
                     /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(4682..4712)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4727..4748
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5036..5624)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5795..6655)
                     /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(6656..6760)
                     /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 ccaccatgga cgagaaggtg
      721 ttcaccaagg agctggacca gtggatcgag cagctgaacg agtgcaagca gctgtccgag
      781 tcccaggtca agagcctctg cgagaaggct aaagaaatcc tgacaaaaga atccaacgtg
      841 caagaggttc gatgtccagt tactgtctgt ggagatgtgc atgggcaatt tcatgatctc
      901 atggaactgt ttagaattgg tggcaaatca ccagatacaa attacttgtt tatgggagat
      961 tatgttgaca gaggatatta ttcagttgaa acagttacac tgcttgtagc tcttaaggtt
     1021 cgttaccgtg aacgcatcac cattcttcga gggaatcatg agagcagaca gatcacacaa
     1081 gtttatggtt tctatgatga atgtttaaga aaatatggaa atgcaaatgt ttggaaatat
     1141 tttacagatc tttttgacta tcttcctctc actgccttgg tggatgggca gatcttctgt
     1201 ctacatggtg gtctctcgcc atctatagat acactggatc atatcagagc acttgatcgc
     1261 ctacaagaag ttccccatga gggtccaatg tgtgacttgc tgtggtcaga tccagatgac
     1321 cgtggtggtt ggggtatatc tcctcgagga gctggttaca cctttgggca agatatttct
     1381 gagacattta atcatgccaa tggcctcacg ttggtgtcta gagctcacca gctagtgatg
     1441 gagggatata actggtgcca tgaccggaat gtagtaacga ttttcagtgc tccaaactat
     1501 tgttatcgtt gtggtaacca agctgcaatc atggaacttg acgatactct aaaatactct
     1561 ttcttgcagt ttgacccagc acctcgtaga ggcgagccac atgttactcg tcgtacccca
     1621 gactacttcc tgctcgagga acaaaaacta atatcagagg aagatttgga acagaaacta
     1681 atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata gtgagggccc
     1741 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1801 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1861 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     1921 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     1981 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2041 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2101 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     2161 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2221 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2281 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2341 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     2401 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     2461 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     2521 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     2581 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     2641 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     2821 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2881 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2941 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3001 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     3061 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3241 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3301 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3361 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3421 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3481 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3541 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3601 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3661 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3721 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3781 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3841 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3901 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3961 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4021 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4081 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4141 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4201 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4261 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     4321 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4381 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4441 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4501 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4561 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4621 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4681 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4741 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4801 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4861 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5221 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6781 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     6841 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     6901 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     6961 gaagaatctg cttagg
//
