LOCUS       V035879                 7432 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035879
VERSION     V035879
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..2237
                     /label="VPS4A(NM_013245)"
                     /note="VPS4A(NM_013245)"
                     /gene="VPS4A"
     CDS             2275..2994
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    3032..3256
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3302..3730
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3744..4073
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4140..4934
                     /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    5108..5229
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5278..5294)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5302..5318
                     /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(5326..5356)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5371..5392
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5680..6265)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6436..7296)
                     /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(7297..7401)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatga caacgtcaac cctccagaaa gccattgatc
      961 tggtgacgaa agccacagag gaggacaaag ccaagaacta cgaggaggcg ctgcggctgt
     1021 accagcatgc ggtggagtac ttcctccacg ctatcaagta tgaggcccac agcgacaagg
     1081 ccaaggagag cattcgagcc aagtgcgtgc agtacctaga ccgggccgag aagctgaagg
     1141 attatttacg aagcaaagag aaacacggca agaagccagt caaagagaac cagagtgagg
     1201 gcaagggcag tgacagtgac agtgaagggg ataatccgga gaaaaagaaa ctgcaagaac
     1261 agctgatggg tgccgtcgtg atggagaagc ccaacatacg gtggaacgac gtggccgggc
     1321 tggagggggc caaggaggcc ctcaaagaag ctgtcatttt gccaatcaaa ttcccacact
     1381 tgttcacagg caagcgcacc ccctggcggg ggattctgct gttcggaccc cctggcacag
     1441 ggaaatccta cctggccaaa gccgtggcaa cagaggccaa caactccacc ttcttctctg
     1501 tgtcctcctc agatctgatg tccaagtggc tgggggagag tgaaaagctg gtcaagaacc
     1561 tgtttgagct ggccaggcag cacaagccct ccatcatctt catcgatgag gtggattccc
     1621 tctgcgggtc ccgaaatgaa aatgagagtg aggccgcccg gaggatcaaa acggagttct
     1681 tggtccagat gcagggggtg gggaataaca atgatgggac tctggttctt ggagccacaa
     1741 acatcccatg ggtgttggat tcggccatca ggaggaggtt tgaaaaacga atttatatcc
     1801 ccttgccgga ggaagctgcc cgcgcccaga tgttccggtt gcatctcggg agcactcccc
     1861 acaacctcac ggatgcaaac atccacgagc tggcccggaa gacggaaggc tactcgggcg
     1921 cggacatcag catcatcgtg cgggactctc tcatgcagcc cgtgaggaag gtgcagtcgg
     1981 ccacacactt caaaaaggtc tgtggcccct ctcgcaccaa ccccagcatg atgattgatg
     2041 acctcctgac tccatgctca ccaggggacc caggagccat ggagatgact tggatggatg
     2101 tccctgggga caaactctta gagcctgtgg tttgcatgtc ggacatgctg cggtctctgg
     2161 ccaccacccg gcccacggtg aatgcagacg acctcctgaa agtgaagaaa ttctcagagg
     2221 actttgggca agagagtctc gagtggcggt ggaggaagcg gcggaggtgg atctatggtg
     2281 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2341 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2401 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2461 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2521 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2581 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2641 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2701 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2761 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2821 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2881 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2941 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtagtctaga
     3001 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3061 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3121 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3181 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3241 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3301 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3361 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3481 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3601 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3661 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3721 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3901 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3961 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4021 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4081 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4141 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4201 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4261 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4321 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4381 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4441 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4501 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4561 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4621 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4681 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4741 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4801 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4861 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4921 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4981 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5041 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5101 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5161 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5221 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5281 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5341 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5401 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
