LOCUS       V038953                 6748 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038953
VERSION     V038953
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"
     CDS             706..1404
                     /label="Rab20(NM_011227)"
                     /note="Rab20(NM_011227)"
                     /gene="Rab20"
     CDS             1411..1500
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1524..2112
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2178..2226
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2428..2856
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2870..3199
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3266..4060
                     /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    4236..4357
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4406..4422)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4430..4446
                     /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(4454..4484)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4499..4520
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4808..5396)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5567..6427)
                     /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(6428..6532)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgcg gaagccggat
      721 gggaagatcg tactgctggg ggacatgaac gtgggtaaaa cgtcgctgct gcagcgctac
      781 atggagcgtc gcttcccgga cacggtcagc accgtgggcg gcgccttcta cctgaagcag
      841 tggcgttcct tcaacatctc catatgggac accgcagggc gggagcagtt tcatggtctg
      901 ggctccctgt actgccgcgg agcggccgct atcatcctta catacgatgt gaaccaccca
      961 cagagcctgt ttgagctgga ggacagattc ctgggcctga cagaaacagc caacaatgac
     1021 tgcctgtttg ccattgtggg gaacaaagtg gacctgacct cggagcggga caccgagggc
     1081 ggggagaagg aaggaccggc ctctggcaag gtgggcagct gtgtctccac caaggtaccc
     1141 aagcaggtgc agccagagga tgcagtggcc ctttacaaga agatcctgaa gtacaagatg
     1201 ctggatgaga gggagatgcc agctgctgag cagatgtgct tcgagaccag tgccaagact
     1261 ggatacaatg tggacctcct ctttgaaacc ttgtttgacc tggtggtacc tatgatcatg
     1321 cggcagaggg ctgaggagtc agatcagact gtggatatag ccagctgcaa aacacccaaa
     1381 cagactagat ccgggtgttg tgccctcgag gaacaaaaac taatatcaga ggaagatttg
     1441 gaacagaaac taatcagtga agaagattta gagcaaaagt taatttccga ggaggactta
     1501 tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     1561 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     1621 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     1681 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     1741 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     1801 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     1861 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     1921 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     1981 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2041 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2101 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     2161 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     2221 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     2281 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     2341 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     2401 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2461 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2521 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2581 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2641 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2701 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2761 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     2821 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2881 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2941 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3001 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3061 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3121 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3181 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3241 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3301 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3361 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3421 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3481 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3541 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3601 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3661 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3721 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3781 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3841 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3901 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3961 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4021 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     4081 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4141 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4201 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4261 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4321 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4381 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4441 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4501 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4561 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4621 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4681 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4741 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4801 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4861 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4921 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4981 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     5041 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5101 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5161 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5221 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5281 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5341 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5401 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5461 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5521 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5581 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5641 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5701 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5761 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5821 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5881 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5941 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6001 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6061 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6121 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6181 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6241 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6301 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6361 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6421 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6481 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6541 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     6601 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6661 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6721 cgacaattgc atgaagaatc tgcttagg
//
