LOCUS       V047207                 8475 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047207
VERSION     V047207
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..3155
                     /label="Dpp10(NM_199021)"
                     /note="Dpp10(NM_199021)"
                     /gene="Dpp10"
     CDS             3162..3227
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3251..3839
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3905..3953
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4155..4583
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4597..4926
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4993..5787
                     /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    5963..6084
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6133..6149)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6157..6173
                     /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(6181..6211)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6226..6247
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6535..7123)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7294..8154)
                     /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(8155..8259)
                     /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 ccaccatgac agccatgaag caggagcagc
      781 aacccacccc aggggccagg gcaacccagt cgcagccagc cgaccaggaa ctaggaagta
      841 acagccctcc acaaagaaac tggaagggaa tcgccattgc cctgctggtg atcttggtgg
      901 tatgctcact catcacaatg tctgtcatcc tgttaacacc agatgaacta acaaattctt
      961 cagaaacaag actgtcacta gaggagcttt tggggaaagg attcggactt cataatccag
     1021 aacctcggtg gatcaatgat acagtcgtgg tatataaaac caacaatgga cacgtcatga
     1081 aactgaatac agaatcaaat gcttccacat tgttattgga caactcaact tttgtaacgt
     1141 ttaaagcctc cagacactca ctttccccag atttaaagta tgttctcctg gcatacgacg
     1201 ttaagcagat ttttcattac tcatttaccg cttcatattt gatttacaac atacacactg
     1261 gggaagtgtg ggagttgaat cccccagaag ttgaggactc agtgttgcag tatgcggcct
     1321 ggggtgtgca aggacagcag ctgatttata tttttgaaaa caatatctac tatcaacctg
     1381 atatcaagag cagttcttta cgactaacat cttcaggaaa ggaaggcatt atttttaatg
     1441 ggattgctga ctggttatat gaagaagaac ttctgcattc tcatatcgcc cactggtggt
     1501 cccccgatgg agagagactt gccttcctga tgataaatga ctcgttggtg cctaacatga
     1561 tcatacctcg gtttactgga gcactgtacc ccaaagcaaa gcagtaccca taccccaagg
     1621 caggtcaagc gaacccatca gtgaagttat atgttgtaaa cctgtatgga ccaactcata
     1681 ctttggagct gatgccaccc gacattttta aatcaagaga atattacatc accatggtta
     1741 aatgggtgag caacacgagg acagtggtca ggtggttgaa tcggcctcag aacatctcca
     1801 tcctcacact ctgtgaatct accaccgggg catgcagcag gaaatatgag atgacgtccg
     1861 acacctggct ctctaaacag aacgaggagc cagtgttttc cagagatgga agcaagttct
     1921 tcatgactgt tcctgttaaa caaggtggaa gaggagaatt ccaccacata gctatgttcc
     1981 tggtccagag taaaagcgag caaattacgg tgcggcatct gacatcagga aactgggaag
     2041 tgataaggat cttggcctat gatgaaacaa ctcaaaaaat ttactttctg agcacagaat
     2101 cttctcccca agggaggcaa ctatacagtg cttctactga gggactattg aatcgtgatt
     2161 gcatctcatg caactttatg aaagaagatt gcacgtattt tgatgccagc tttagcccca
     2221 tgaatcaaca tttcttgtta ttctgtgaag gtccaaaggt cccagtggtc agccttcaca
     2281 tcacagacaa cccatcaagg tatttcctct tggaaaacaa ttctgtgatg aaagaaacta
     2341 ttcagaagaa gaagctcgca aagagagaga ctagaatact tcacattgat gactatgaac
     2401 ttcctttaca gttgtccttt cccaaagatt ttatggagaa aaaccagtat gctcttctat
     2461 taataatgga tgaagaacca ggaggccaaa tggtgacaga taagttccat gttgactggg
     2521 attcagttct tattgacacc gataatgtca ttgtagcaag atttgatggc agaggaagtg
     2581 gattccaggg cctgaaagtt ttgcaggaga ttcacagaag gataggctca gtggaggcaa
     2641 aggaccaagt agctgctgta aaatatttac tgaaacagcc atatattgac tccaaaagat
     2701 taagcatttt tggaaaggga tatgggggct atattgcatc aatgatctta aaatcagatg
     2761 agaagttttt caaatgtgga gccgtggttg cacccatctc agacatgaag ttgtatgcct
     2821 cagctttctc tgaacggtac cttggcatgc catcaaagga agaaagcact taccaggcat
     2881 ccagtgtact gcataacatt catggtttaa aagaagaaaa tttattaata attcacggaa
     2941 ctgctgatac aaaagttcat ttccagcatt cagcagaatt gatcaagcat ctgataaaag
     3001 ctggggtgaa ttacactctg caggtctatc cagatgaagg atatcacatt tcagacaaga
     3061 gcaagcatca tttttacagc acaattctca gattcttcag tgattgtcta aaggaagagg
     3121 tatctgtgct gccacaggaa ccagaagaag atgaaggatc cgactacaaa gaccatgacg
     3181 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     3241 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3301 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3361 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3421 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3481 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3541 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3601 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3661 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3721 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3781 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3841 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3901 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     3961 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4021 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4081 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     4141 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4201 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4261 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     4321 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4381 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4441 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4501 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     4561 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4621 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4681 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4741 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4801 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4861 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4921 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5161 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5521 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5821 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5881 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5941 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6061 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6121 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6721 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6961 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8281 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     8341 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     8401 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     8461 aagaatctgc ttagg
//
