LOCUS       V034323                 8478 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034323
VERSION     V034323
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..3158
                     /label="RAB11FIP3(NM_001370401)"
                     /note="RAB11FIP3(NM_001370401)"
                     /gene="RAB11FIP3"
     CDS             3165..3230
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3254..3842
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3908..3956
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4158..4586
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4600..4929
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4996..5790
                     /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    5966..6087
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6136..6152)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6160..6176
                     /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(6184..6214)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6229..6250
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6538..7126)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7297..8157)
                     /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(8158..8262)
                     /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 gtcggccccg ccggcctcgc
      781 ccccgggctc ggagccgccg gggcccgacc cggagccggg cgggccggac gggccggggg
      841 cggcacaact ggctccgggc cctgcggagc tacgcctcgg agcgcccgtc ggcggccccg
      901 acccgcagtc cccgggcctg gatgagcctg cgcccggggc cgctgcagat ggcggggcgc
      961 gttggagcgc cgggccggcc ccggggctgg agggaggccc gcgagacccc gggccgtccg
     1021 ccccgccgcc gcgctccggc ccgcgggggc agcttgcgag ccccgacgcc ccgggcccag
     1081 ggccgcgctc cgaagcgccg cttccagaac tcgacccgtt gttctcctgg actgaggagc
     1141 ccgaggagtg tggccccgcg agctgcccgg agagcgcgcc tttccgcttg caggggtcca
     1201 gcagcagcca ccgagcgcgg ggcgaggtcg acgtcttctc tcccttcccc gcgcccacgg
     1261 cgggcgagct ggcgctggag caaggtcccg ggtccccgcc gcagccctcg gacctcagcc
     1321 agacccaccc ccttccgagc gagcccgtgg ggagtcagga ggacggcccc cgcctccgag
     1381 ccgtgttcga tgccctggac ggggatgggg acggtttcgt ccgcatcgag gacttcatcc
     1441 agtttgctac ggtctacggg gcagagcagg tgaaggactt aactaagtac ttggatccca
     1501 gtgggctcgg cgtgatcagc tttgaagact tctaccaagg gatcacagcc atcagaaacg
     1561 gagatcctga tggccagtgc tacggtggtg tcgcttctgc ccaagatgag gagcccctgg
     1621 cctgcccgga cgagttcgat gacttcgtca cctatgaggc caacgaggtg acggacagcg
     1681 cgtacatggg ctccgagagc acctacagtg agtgtgagac cttcacggac gaggacacca
     1741 gcaccctggt gcaccctgag ctgcaacctg aaggggacgc agacagtgcc ggcggctcgg
     1801 ccgtgccctc tgagtgcctg gacgccatgg aggagcccga ccatggtgcc ctgctgctgc
     1861 tcccaggcag gcctcacccc catggccagt ctgtcatcac ggtgatcggg ggcgaggagc
     1921 actttgagga ctacggtgaa ggcagtgagg cggagctgtc cccagagacc ctatgcaacg
     1981 ggcagctggg ctgcagtgac cccgctttcc tcacgcccag cactgaccct cttgccgcaa
     2041 agctgcacag catcctcact gatgaggcgt ttgagtttta ctgtagccag tgccacaaac
     2101 aaatcaaccg ccttgaggat ctttctgccc gcctgagtga tcttgaaatg aatagtccga
     2161 caaagcggct ctccagcaag aaggtggcaa ggtacctgca ccagtcaggg gccctgacca
     2221 tggaggccct ggaggaccct tcccccgagc tcatggaggg cccagaggag gacattgctg
     2281 acaaggttgt cttcctggaa aggcgtgtgc tggagctgga aaaggacacg gcagccaccg
     2341 gtgagcaaca cagccgcctg aggcaggaga acctgcagct ggtgcacaga gcaaacgccc
     2401 tggaggagca gctgaaggag caggagctga gagcctgcga gatggtcctg gaagagaccc
     2461 ggcgtcagaa ggagctcctg tgcaagatgg agagggagaa gagcattgag atcgagaacc
     2521 tgcagaccag gctacagcaa ctggacgagg agaacagtga actccggtcc tgcacgccct
     2581 gtctgaaggc caacattgag cgtctggagg aggagaagca gaagctgttg gatgagatag
     2641 agtcgctgac gctgcggctc agtgaagagc aggagaacaa gaggagaatg ggggacaggc
     2701 tgagtcacga gaggcaccag ttccagaggg acaaggaggc cacccaggag ctgatcgagg
     2761 acctccgaaa gcagctggag cacctgcagc tcctcaagct ggaggccgag cagcggcggg
     2821 gccgcagcag cagcatgggc ctgcaggagt accacagccg cgcccgggag agcgagctgg
     2881 agcaggaggt ccgcaggctg aagcaggaca accgcaacct gaaggagcag aacgaggagc
     2941 tgaacgggca gatcattacc ctcagcatcc agggcgccaa gagcctcttc tccacagcct
     3001 tctctgagtc cctggctgca gagatcagct ccgtctcccg agatgagctc atggaggcga
     3061 ttcagaagca ggaggagatc aacttccgcc tgcaggacta catcgacagg atcatcgtgg
     3121 ccatcatgga gaccaacccg tccatcctgg aggtcaaggg atccgactac aaagaccatg
     3181 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     3241 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3301 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3361 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3421 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3481 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3541 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3601 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3661 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3721 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3781 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3841 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3901 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3961 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4021 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4081 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4141 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4201 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4261 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     4321 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4381 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4441 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4501 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4561 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4621 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4681 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4741 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4801 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4861 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4921 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4981 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5041 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5101 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5161 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5221 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5281 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5341 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5401 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5461 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5521 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5581 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5641 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5701 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5761 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     5821 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5881 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5941 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6001 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6061 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6121 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6181 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6241 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6301 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6361 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6421 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6481 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6541 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6601 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6661 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6721 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6781 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6841 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6901 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6961 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7021 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7081 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7141 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7201 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7261 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7321 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7381 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7441 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7501 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7561 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7621 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7681 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7741 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7801 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7861 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7921 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7981 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8041 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8101 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8161 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8221 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8281 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     8341 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8401 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8461 atgaagaatc tgcttagg
//
