LOCUS       V036698                10236 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036698
VERSION     V036698
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..4916
                     /label="Rab3gap2(NM_001163754)"
                     /note="Rab3gap2(NM_001163754)"
                     /gene="Rab3gap2"
     CDS             4923..4988
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5012..5600
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5666..5714
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5916..6344
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6358..6687
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6754..7548
                     /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    7724..7845
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7894..7910)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7918..7934
                     /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(7942..7972)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7987..8008
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8296..8884)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9055..9915)
                     /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(9916..10020)
                     /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 ccaccatggc ctgctccatc gtccagttct
      781 gctccttcca ggacctgcag tccgcgcgcg acttcctatt ccctcacctg cgggaggaga
      841 cccccggcgc cctgaagagg gacccgagta aaacatcaag ctgggaagat gatagttggg
      901 gagcatggga agagacagag ccccgagaac ctgaagaaga aggaaatact tctaaaacac
      961 aaaaaaattc ctggcttcaa gaatgtgttt tatccttatc tccaaccagt gaccttatgg
     1021 ttatagctcg ggagcagaag gccgcattcc tagtgccaaa atggaaacat ggtgataaag
     1081 ggaaggaaga aatgcagttt gctgttggtt ggagcggctc cgtaagtgca gaggaagggg
     1141 agtatgtaac cagtgccctg tgcatcccac tggcgagtca gaagaggagt tccactgggc
     1201 gccctgactg gacctgcatt gtggtgggct tcacatcggg ctatgtgcgc ttctacactg
     1261 agagtggagt gcttttgctt gcacaacttt tgaatgagga caaagtgctt cagcttaaat
     1321 gcaggaccta tgaaatccca agacaccctg gagtgactga gcagaatgaa gaactgagca
     1381 tcttatatcc agccgccatt gtgacgattg atggatttag tctctttcag tctcttcgag
     1441 cctgtcgaaa ccaagtagca aaagctgcag catcaggcaa tgagaacatt cagccaccac
     1501 cattagctta taagaaatgg ggtcttcaag atattgacac catcatcgat catgctagta
     1561 ttggtattat gactctgtca ccatttgatc aaatgaagac tgcttccaat ataggtggat
     1621 ttaatgcagc aattaaaaac agtccgcctg ccatgtctca gtacatcact gtgggctcca
     1681 gtccgttcac tggcttcttc tatgctctcg agggaagcac gcagcctctg ttgtctcacg
     1741 tcgcgctcgc tgttgcaagt aagctgacct ccgccctctt cagtgccgcc agtggttggc
     1801 ttggttggaa aagcaagcat gaggaagaca ctgtccagaa gcagaagccc aagatggagc
     1861 cggccacccc actagctgtg agatttggac ttccggattc tagacgccac ggagaaagca
     1921 tatgcctgtc tccttgtaat accctggcag cagtcacaga tgattttggc agggttatct
     1981 tactggacgt agcccgagga atcgcaatac ggatgtggaa agggtatcgg gatgcgcagg
     2041 tcggatggat tcaaattgta gaggacctcc atgaacgagt gccagaaaag gggggcttct
     2101 ccccctttgg aaatactcaa gggcccagtc gggtcgctca gttccttgtg atctatgcac
     2161 cacgaagggg gattctggaa gtgtggaaca cacagcaagg acctagagtg ggagctttca
     2221 acgtgggtaa acactgcagg ctactctatc ctggctacaa aatcatgggc ttaaacaacg
     2281 tgacaagtca gagttggcag cctcagactt accagatctg tcttgttgac cctgtgtctg
     2341 caagcgtgaa ggcggtgaac gtgcctttcc atctagcgct gagtgataag aagagtgaaa
     2401 gagccaagga cctgcactta gtgaagaaac tatcagccct gctgagagcc aagtccccca
     2461 ggccggattc atttgaaacc gaaataaagg agttgattct tgatattaag taccctgcaa
     2521 ccaagaaaca agctctggaa agcattttgg ccagtgaccg tttatcgttt tcctgcctta
     2581 gaaatgttac tcagacttta atggacactt taaaaaacca agagcttgag tctgttgatg
     2641 aaggactgct acagttttgt gccagtaaat taaaactact acatctttat gaatctgtca
     2701 gtcagttaaa tacgcttgac tttcactcag acaccccgtt ctctgataat gatctggctg
     2761 tcttgctaag gctggatgat aaggaactcc tgaaactccg ggccttgtta gagaaatata
     2821 aacaagaaaa caccaaagcc accgtgcgct tttctgagga tgcagacaga gtgctacctg
     2881 tcaagacctt cctggagtat ctagagtatg agaaggatgc actgagcata aggaaggtcg
     2941 gagaggagga gtgtgtggct ctaggcagtt tctttttctg gaagtgtttg catggagaaa
     3001 gctccactga agagatgtgc cacagcctgg agtcggctgg actcagccct cagcagttac
     3061 tgtctctgct cctaagtgtt tggctttcga aggaaaaaga cattttggac aaaccacagt
     3121 ctgtctgctg ccttcatacc atgctgtccc tgctgagcaa gatgaaagtg gccatcgatg
     3181 agacgtggga ttctcagtct gtgtccccct ggtggcagca gatgcgcatg gcctgcatcc
     3241 agtcagagaa cagcggcgct gccctgctgt ctgcacatgt ggggcattct gtggccgcac
     3301 agatgtccag cggtgctaca gataagaaat tttcccagat ggtgttggat gcagatgccg
     3361 aagccctcac ggactcctgg gaagctctct ctctggacac ggagtactgg aagctcctgc
     3421 tgagacagct ggaggactgc ctcatactcc agactctgct gcatagcaaa ttgagccccc
     3481 cagccgccaa agcaccatct ctgcagtcgg agccactgcc aaggctttct gttaaaaagc
     3541 tgttagaagg ggggaaaggt ggtattgcag acagtgtagc caagtggata tttaaacagg
     3601 acctcagccc tgaattgttg aaatgtgcca acaaagagag agatgtagaa aatccagacg
     3661 aacccagaga aggtattaca cgaagtttcc ctgaggtgtc agaggtggag atagacctag
     3721 gagccgtacc agacttgctc cacctggcct acgagcagtt cccttgtagc cttgagcttg
     3781 atgtgctgca cgcccactgc tgctgggagt atgtggttca gtggaacaaa gacccggagg
     3841 aagctcgctt tttggttaga tccatagaac acttgaagca cattcttaat ccacacgttc
     3901 aaaatggtat cgcgctgatg atgtggaata cgttcttggt gaaaaggttt tctgctgcta
     3961 cttacctaat ggataaggtt gggaaatctc caaaggatag gttgtgccga agggatgtgg
     4021 ggatgagcga cacggcgctg acatccttcc tcggctcctg cctggaactt cttcagacgt
     4081 cattggaggc tgacattagc agggatgaag tccaggtgcc cgtcctggac actgaggacg
     4141 cctggctgtc cgtggaaggc cccatctcta tagtggagct ggcgctggag cagaagccta
     4201 tccactaccc actggtggag caccattccg tcctgtgctc catcctgtac gcgtccatgc
     4261 gcttttctct gaagagtgtg aagccactcg ccctttttga cagcaaggga aagaatgcat
     4321 tttttaagga tttaacttca attcagttat tacctagtgg ggaaatggat ccaaatttta
     4381 tttcagtaag acaacagttc ttactgaaag ttgtcagtgc agctgtccag gctcagcatt
     4441 ccaaggacaa agatccctct gcagaagcag caaacactca ttggaaagac ctcaactggc
     4501 caggtttggc tgtggactta gcccatcacc ttcaagtgag cgacgatgtc attcgaaggc
     4561 actatgtggg ggagctgtac agccatggag ctgacctctt aggagaggag gccatttttc
     4621 aagttcagga caaagaagtt ctggcgtctc agctgctagt gctgaccggg caaaggctgg
     4681 ctcacgcgct tttccacacc cagacgaagg aaggcatgga gctgctggcc agactcccgc
     4741 ccacactctg cacctggctg aaggcaatga accctcagga tcttcagaac actggcgtgc
     4801 ctattgcagc aacagctaaa ctcgtgcata aagtgatgga gctcctgcca gagaagcacg
     4861 ggcagtacag cctcgcctta cacctcatcg atgccgtgga agccatggcg actttaggat
     4921 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     4981 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     5041 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5101 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5161 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5221 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5281 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5341 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5401 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5461 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5521 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5581 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     5641 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     5701 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5761 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5821 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5881 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     5941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6061 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6121 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6241 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     6301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     6361 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     6481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     6541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6601 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     6661 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     6721 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6781 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6841 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6901 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     6961 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     7021 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     7081 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     7141 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     7201 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     7261 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     7321 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7381 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7441 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7501 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7561 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     7621 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     7681 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     7741 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     7801 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     7861 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     7921 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     7981 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     8041 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     8101 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     8161 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     8221 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     8281 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8341 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8401 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8461 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     8521 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8581 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8641 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8701 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     8761 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8821 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8881 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8941 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     9001 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9061 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9121 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9181 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9241 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9301 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9361 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9421 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9481 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9541 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9601 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9661 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9721 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9781 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9841 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9901 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9961 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    10021 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
    10081 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
    10141 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
    10201 ggcttgaccg acaattgcat gaagaatctg cttagg
//
