LOCUS       V020144                 9224 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020144
VERSION     V020144
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             683..3904
                     /label="PLD1(NM_002662)"
                     /note="PLD1(NM_002662)"
                     /gene="PLD1"
     CDS             3911..3976
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4000..4588
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4654..4702
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4904..5332
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5346..5675
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5742..6536
                     /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    6712..6833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6882..6898)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6906..6922
                     /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(6930..6960)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6975..6996
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7284..7872)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8043..8903)
                     /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(8904..9008)
                     /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 ggatccgcca ccatgtcact gaaaaacgag ccacgggtaa atacctctgc
      721 actgcagaaa attgctgctg acatgagtaa tatcatagaa aatctggaca cgcgggaact
      781 ccactttgag ggagaggagg tagactacga cgtgtctccc agcgatccca agatacaaga
      841 agtgtatatc cctttctctg ctatttataa cactcaagga tttaaggagc ctaatataca
      901 gacgtatctc tccggctgtc caataaaagc acaagttctg gaagtggaac gcttcacatc
      961 tacaacaagg gtaccaagta ttaatcttta cactattgaa ttaacacatg gggaatttaa
     1021 atggcaagtt aagaggaaat tcaagcattt tcaagaattt cacagagagc tgctcaagta
     1081 caaagccttt atccgcatcc ccattcccac tagaagacac acgtttagga ggcaaaacgt
     1141 cagagaggag cctcgagaga tgcccagttt gccccgttca tctgaaaaca tgataagaga
     1201 agaacaattc cttggtagaa gaaaacaact ggaagattac ttgacaaaga tactaaaaat
     1261 gcccatgtat agaaactatc atgccacaac agagtttctt gatataagcc agctgtcttt
     1321 catccatgat ttgggaccaa agggcataga aggtatgata atgaaaagat ctggaggaca
     1381 cagaatacca ggcttgaatt gctgtggtca gggaagagcc tgctacagat ggtcaaaaag
     1441 atggttaata gtgaaagatt cctttttatt gtatatgaaa ccagacagcg gtgccattgc
     1501 cttcgtcctg ctggtagaca aagaattcaa aattaaggtg gggaagaagg agacagaaac
     1561 gaaatatgga atccgaattg ataatctttc aaggacactt attttaaaat gcaacagcta
     1621 tagacatgct cggtggtggg gaggggctat agaagaattc atccagaaac atggcaccaa
     1681 ctttctcaaa gatcatcgat ttgggtcata tgctgctatc caagagaatg ctttagctaa
     1741 atggtatgtt aatgccaaag gatattttga agatgtggca aatgcaatgg aagaggcaaa
     1801 tgaagagatt tttatcacag actggtggct gagtccagaa atcttcctga aacgcccagt
     1861 ggttgaggga aatcgttgga ggttggactg cattcttaaa cgaaaagcac aacaaggagt
     1921 gaggatcttc ataatgctct acaaagaggt ggaactcgct cttggcatca atagtgaata
     1981 caccaagagg actttgatgc gtctacatcc caacataaag gtgatgagac acccggatca
     2041 tgtgtcatcc accgtctatt tgtgggctca ccatgagaag cttgtcatca ttgaccaatc
     2101 ggtggccttt gtgggaggga ttgacctggc ctatggaagg tgggacgaca atgagcacag
     2161 actcacagac gtgggcagtg tgaagcgggt cacttcagga ccgtctctgg gttccctccc
     2221 acctgccgca atggagtcta tggaatcctt aagactcaaa gataaaaatg agcctgttca
     2281 aaacctaccc atccagaaga gtattgatga tgtggattca aaactgaaag gaataggaaa
     2341 gccaagaaag ttctccaaat ttagtctcta caagcagctc cacaggcacc acctgcacga
     2401 cgcagatagc atcagcagca ttgacagcac ctccagttat tttaatcact atagaagtca
     2461 tcacaattta atccatggtt taaaacccca cttcaaactc tttcacccgt ccagtgagtc
     2521 tgagcaagga ctcactagac ctcatgctga taccgggtcc atccgtagtt tacagacagg
     2581 tgtgggagag ctgcatgggg aaaccagatt ctggcatgga aaggactact gcaatttcgt
     2641 cttcaaagac tgggttcaac ttgataaacc ttttgctgat ttcattgaca ggtactccac
     2701 gccccggatg ccctggcatg acattgcctc tgcagtccac gggaaggcgg ctcgtgatgt
     2761 ggcacgtcac ttcatccagc gctggaactt cacaaaaatt atgaaatcaa aatatcggtc
     2821 cctttcttat ccttttctgc ttccaaagtc tcaaacaaca gcccatgagt tgagatatca
     2881 agtgcctggg tctgtccatg ctaacgtaca gttgctccgc tctgctgctg attggtctgc
     2941 tggtataaag taccatgaag agtccatcca cgccgcttac gtccatgtga tagagaacag
     3001 caggcactat atctatatcg aaaaccagtt tttcataagc tgtgctgatg acaaagttgt
     3061 gttcaacaag ataggcgatg ccattgccca gaggatcctg aaagctcaca gggaaaacca
     3121 gaaataccgg gtatatgtcg tgataccact tctgccaggg ttcgaaggag acatttcaac
     3181 cggcggagga aatgctctac aggcaatcat gcacttcaac tacagaacca tgtgcagagg
     3241 agaaaattcc atccttggac agttaaaagc agagcttggt aatcagtgga taaattacat
     3301 atcattctgt ggtcttagaa cacatgcaga gctcgaagga aacctagtaa ctgagcttat
     3361 ctatgtccac agcaagttgt taattgctga tgataacact gttattattg gctctgccaa
     3421 cataaatgac cgcagcatgc tgggaaagcg tgacagtgaa atggctgtca ttgtgcaaga
     3481 tacagagact gttccttcag taatggatgg aaaagagtac caagctggcc ggtttgcccg
     3541 aggacttcgg ctacagtgct ttagggttgt ccttggctat cttgatgacc caagtgagga
     3601 cattcaggat ccagtgagtg acaaattctt caaggaggtg tgggtttcaa cagcagctcg
     3661 aaatgctaca atttatgaca aggttttccg gtgccttccc aatgatgaag tacacaattt
     3721 aattcagctg agagacttta taaacaagcc cgtattagct aaggaagatc ccattcgagc
     3781 tgaggaggaa ctgaagaaga tccgtggatt tttggtgcaa ttcccctttt atttcttgtc
     3841 tgaagaaagc ctactgcctt ctgttgggac caaagaggcc atagtgccca tggaggtttg
     3901 gactctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     3961 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     4021 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4081 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4141 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4201 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4261 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4321 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4381 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4441 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4501 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4561 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     4621 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4681 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4741 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4801 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4861 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4981 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5041 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     5101 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5221 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     5281 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5341 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5401 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5461 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5581 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5641 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5701 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5761 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5821 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5881 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5941 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6001 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6061 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6121 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6181 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6241 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6301 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6361 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6421 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6481 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6541 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6601 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6661 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6841 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6901 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6961 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7021 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7441 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7681 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7921 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7981 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8041 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8101 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8161 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8221 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8281 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8341 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8401 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8461 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8521 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8581 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8641 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8701 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8761 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8821 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8881 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8941 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9001 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     9061 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9121 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9181 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
