LOCUS       V044096                 9078 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044096
VERSION     V044096
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             756..3758
                     /label="Ptprn2(NM_011215)"
                     /note="Ptprn2(NM_011215)"
                     /gene="Ptprn2"
     CDS             3765..3830
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3854..4442
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4508..4556
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4758..5186
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5200..5529
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5596..6390
                     /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    6566..6687
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6736..6752)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6760..6776
                     /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(6784..6814)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6829..6850
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7138..7726)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7897..8757)
                     /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(8758..8862)
                     /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 ctcgaattcg ccaccatggg gccgccgctc ccgcttttgc
      781 tgctgctgct gctaccgccg ccgctgcctc gcgctctgcc cgcccccgcg tctgcccgcg
      841 gccggcagct cccggggcgc ctgggatgct tgtttgagga tggcttgtgt ggatcactgg
      901 agacctgtgt gaacgatggt gtgtttggaa gatgtcaaaa ggttccggtg atggacactt
      961 accgatatga ggtaccacca ggagccctgc tgcacctgaa ggtcacctta cagaagctct
     1021 cccgtacagg tttcacgtgg caagatgact atacccagcg tgtgattgcc caggagcttg
     1081 caaacctccc caaggcctac ctatggcatg gggaagcgtc cggcccagcc aggtccttac
     1141 aacagaatgc tgacaatgaa aaatggttca gtctggagag ggaggtggcc ctggccaaga
     1201 cccttcggcg ctatctgccc tacctggagc ttctgtccca gaccccaaca gcaaatgcac
     1261 actctaggat agaccatgag actcgtccag ccaagggtga agactcttcc cctgagaaca
     1321 tcctgaccta cgtggcccac acatcagcac tgacctatcc tcctgcaacc cgggccaagt
     1381 atcctgataa ccttctgcgg ccctttagcc ggctccagcc agatgagctc agccccaagg
     1441 tagacggtga catagacaaa cagaagctga ttgcagcact gggcgcctac actgctcaga
     1501 ggcttcctgg agaaaatgac ccagagccac ggtaccttgt acatggttcc gcgagagcac
     1561 caaggccatt ctcagcaact gctttgtctc agagatggcc tccacctcct ggagacgcca
     1621 aagactcccc gagtatggat gatgacacac tcctgcagag tctcctgaag gatttgcagc
     1681 agaactctga agtggaccgc ctgggccccc tgaaggagga gaaagcagac tcagttgctg
     1741 gagccataca aagtgatcct gcagagggaa gccaagaaag ccacgggaga ggggctgaag
     1801 gacagccaag agagcagaca gatgccccag agacaatgct tcaagatcac agactatcag
     1861 aggtggatga cccagtgtac aaggaggtca accgtctgag cttccagctt ggggacctct
     1921 tgaaggacta tgggtctcct ctcttacctg aaggtcccct tctagaaaaa tcctccagag
     1981 aagagattaa gaaatcagag cagccagagg aggtcttgtc ttcagaagag gagactgctg
     2041 gggtggagca tgtgaggagc cggacttact ccaaagacct atttgaaagg aaaccaaact
     2101 cagagcccca gcccaggagg cttgaggatc agttccaaaa ccgagctcca gagttgtggg
     2161 aggatgaaga aagcctcaaa ttggcagcac agggaccccc tagtggaggc ctacagctgg
     2221 aagtgcagcc ttctgaggaa cagcagggat acatcctcac aggaaacaac cctctaagtc
     2281 cagagaaggg gaagcagctg atggaccaag ttgcccacat cctccgggta ccttccagct
     2341 tctttgcaga tatcaaagtt ttgggaccag cagtgacctt caaagtaagt gccaacatcc
     2401 aaaacatgac aactgccgat gtcatcaagg ctgcagctga caacaaagac cagctggaga
     2461 aggcaactgg actgacaatc cttcaaagtg gaatcaggcc gaagggaaag ctcaaactcc
     2521 tgccgcatca ggaagagcaa gaggactcta ccaagttcat tttgctcacc ttcctctcca
     2581 ttgcctgcat cctgggggtt ctcctggctt ccagccttgc ctactgcctc cgccacaact
     2641 cacactacaa gctgaaggac aagttgtctg gactaggcgc tgaccccagt gcagatgcca
     2701 ctgaagccta ccaggagcta tgccgccagc gtatggctat tcgtccacag gaccgctctg
     2761 agggaccaca tacatcacgc atcaacagcg tctcatccca gttcagcgat gggccgatgc
     2821 ctagtccttc ggctcggagc agcacttcat cctggtctga ggagcctgtc cagtccaaca
     2881 tggacatctc tactggccac atgatcctgg cctacatgga agaccatctg aagaacaaga
     2941 accgcctgga gaaggagtgg gaagcactgt gcgcctacca agcagagccc aacagctcac
     3001 ttgtggccca gagagaggag aatgcaccca agaaccgttc cctggctgtg ctgacctatg
     3061 accactccag gatcctgttg aagtctcaaa acagccatgg cagttccgac tacatcaatg
     3121 ccagccccat tatggaccat gacccacgaa accccgcata cattgccacc caaggcccac
     3181 ttcccgccac ggtggccgac ttctggcaga tggtgtggga aagcggctgt gcagtcattg
     3241 tcatgctgac acccctctcc gagaacggcg tccggcagtg ccatcactac tggcccgatg
     3301 aaggctccaa cctctaccat gtctacgagg tcaatctagt ctctgagcac atatggtgcc
     3361 aggatttcct ggtgagaagc ttttacctga agaacctgca gaccaacgag actcgcacgg
     3421 tgacccagtt ccacttcctg agttggtatg accagggagt cccttcctcc acgaggtcac
     3481 tcctggattt ccgcagaaaa gtgaacaaat gctaccgagg ccgctcttgt ccgatcattg
     3541 tccattgcag tgacggcgcc ggcaggagtg gaacctacgt cctgattgac atggttctca
     3601 ataagatggc caaaggtgct aaagagattg atatcgcagc gaccctggag cacttgaggg
     3661 accagagacc aggcatggtc cagacaaagg agcagtttga gtttgcgctg acagctgtgg
     3721 ctgaggaggt gaatgccatc ctgaaggccc ttccccaggg atccgactac aaagaccatg
     3781 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     3841 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3901 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3961 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4021 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4081 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4141 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4201 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4261 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4321 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4381 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4441 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4501 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4561 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4621 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4681 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4741 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4801 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4861 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     4921 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4981 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5041 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5101 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     5161 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5281 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5341 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5401 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5461 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5521 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5581 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5641 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5701 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5761 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5821 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5881 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5941 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6001 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6061 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6121 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6181 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6241 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6301 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6361 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6421 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6481 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6541 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6601 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6661 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6721 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6781 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6841 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6961 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7021 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7321 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7561 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7741 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7801 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7861 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7921 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7981 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8041 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8101 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8161 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8221 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8281 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8341 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8401 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8461 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8521 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8581 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8641 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8701 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8761 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8821 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8881 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     8941 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     9001 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     9061 atgaagaatc tgcttagg
//
