LOCUS       V024186                 7214 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024186
VERSION     V024186
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             779..1894
                     /label="LPAR5(NM_020400)"
                     /note="LPAR5(NM_020400)"
                     /gene="LPAR5"
     CDS             1901..1966
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1990..2578
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2644..2692
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2894..3322
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3336..3665
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3732..4526
                     /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    4702..4823
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4872..4888)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4896..4912
                     /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(4920..4950)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4965..4986
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5274..5862)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6033..6893)
                     /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(6894..6998)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gttagccaac agctcctcaa ccaacagttc tgttctcccg tgtcctgact accgacctac
      841 ccaccgcctg cacttggtgg tctacagctt ggtgctggct gccgggctcc ccctcaacgc
      901 gctagccctc tgggtcttcc tgcgcgcgct gcgcgtgcac tcggtggtga gcgtgtacat
      961 gtgtaacctg gcggccagcg acctgctctt caccctctcg ctgcccgttc gtctctccta
     1021 ctacgcactg caccactggc ccttccccga cctcctgtgc cagacgacgg gcgccatctt
     1081 ccagatgaac atgtacggca gctgcatctt cctgatgctc atcaacgtgg accgctacgc
     1141 cgccatcgtg cacccgctgc gactgcgcca cctgcggcgg ccccgcgtgg cgcggctgct
     1201 ctgcctgggc gtgtgggcgc tcatcctggt gtttgccgtg cccgccgccc gcgtgcacag
     1261 gccctcgcgt tgccgctacc gggacctcga ggtgcgccta tgcttcgaga gcttcagcga
     1321 cgagctgtgg aaaggcaggc tgctgcccct cgtgctgctg gccgaggcgc tgggcttcct
     1381 gctgcccctg gcggcggtgg tctactcgtc gggccgagtc ttctggacgc tggcgcgccc
     1441 cgacgccacg cagagccagc ggcggcggaa gaccgtgcgc ctcctgctgg ctaacctcgt
     1501 catcttcctg ctgtgcttcg tgccctacaa cagcacgctg gcggtctacg ggctgctgcg
     1561 gagcaagctg gtggcggcca gcgtgcctgc ccgcgatcgc gtgcgcgggg tgctgatggt
     1621 gatggtgctg ctggccggcg ccaactgcgt gctggacccg ctggtgtact actttagcgc
     1681 cgagggcttc cgcaacaccc tgcgcggcct gggcactccg caccgggcca ggacctcggc
     1741 caccaacggg acgcgggcgg cgctcgcgca atccgaaagg tccgccgtca ccaccgacgc
     1801 caccaggccg gatgccgcca gtcaggggct gctccgaccc tccgactccc actctctgtc
     1861 ttccttcaca cagtgtcccc aggattccgc cctcctcgag gactacaaag accatgacgg
     1921 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     1981 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2041 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2101 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2161 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2221 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2281 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2341 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2401 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2461 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2521 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2581 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     2641 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2701 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     2761 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     2821 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     2881 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2941 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3121 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3241 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3301 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3481 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3601 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3661 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3721 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3781 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3841 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3901 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3961 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4021 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4081 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4141 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4201 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4261 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4321 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4381 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4441 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4501 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4561 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4621 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4681 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4741 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4801 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4861 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5041 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5101 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5161 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5221 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5281 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5341 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5401 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5461 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5521 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5581 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5641 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5701 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5761 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5821 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5881 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5941 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6001 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6061 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6121 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6181 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6241 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6301 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6361 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6421 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6481 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6541 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6601 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6661 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6721 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6781 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6841 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6901 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6961 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7021 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7081 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7141 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7201 agaatctgct tagg
//
