LOCUS       V037807                10401 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037807
VERSION     V037807
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             887..5170
                     /label="VPS8(NM_001009921)"
                     /note="VPS8(NM_001009921)"
                     /gene="VPS8"
     CDS             5207..5917
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     polyA_signal    5968..6192
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6238..6666
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6680..7009
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7076..7870
                     /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    8044..8165
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8214..8230)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8238..8254
                     /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(8262..8292)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8307..8328
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8616..9201)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9372..10232)
                     /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(10233..10337)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcaac ttaaggatcc gccaccatgg aaaatgaacc
      901 agaccatgaa aatgtggaac agagcctctg tgccaagacg agcgaagaag agctgaataa
      961 gtctttcaat ctagaagctt cactttcaaa attctcttac atagatatgg acaaggaact
     1021 ggagttcaaa aatgatctga ttgatgacaa ggagtttgat attcctcaag ttgatactcc
     1081 tccaacactg gaaagcatac taaatgagac tgatgatgaa gatgagtctt ttattcttga
     1141 ggatcctaca ttgttaaaca ttgatactat tgattctcac tcctatgata cttcatctgt
     1201 ggcaagctca gatagtggtg acaggaccaa cttaaaaagg aagaagaaat tacctgattc
     1261 tttttcactt catggatcag ttatgcgcca ttcacttttg aagggaattt ctgcccagat
     1321 agtgtctgca gctgacaaag tagatgctgg cttgcctaca gcaattgcag tatccagtct
     1381 gatagcagtg ggtacatctc atggattggc tttaatattt ggaaaagatc agaatcaagc
     1441 tttgcgactc tgtctgggta gcactagtgt tggaggtcag tatggcgcta tctctgccct
     1501 cagtatcaac aatgattgct caagacttct ttgtggcttt gctaaaggac agatcaccat
     1561 gtgggatttg gccagtggaa aacttctaag atcaataaca gatgctcatc ctccaggaac
     1621 agcaatattg catatcaagt ttacagatga tccaactctt gcaatttgca acgacagcgg
     1681 aggctctgtt tttgaattga catttaagag agtgatggga gtgagaacct gtgaatctag
     1741 atgtctgttc agtggttcca agggtgaagt ctgctgtatt gagcctctgc attctaagcc
     1801 tgagttgaaa gatcatccca tcacacagtt ttcattattg gccatggcat ccttgacaaa
     1861 aatactggtc attggattga aaccatcctt gaaagtatgg atgacttttc cctatggccg
     1921 gatggatcct tccagtgtgc cactgctggc ctggcacttt gtagcagtac aaaattacgt
     1981 gaatcccatg cttgccttct gcagaggaga tgttgttcat tttctattgg taaagagaga
     2041 tgaatctgga gcaatacatg ttactaagca aaagcatctt cacctatact atgacctcat
     2101 caactttacc tggataaatt cacgcacagt tgtgctctta gacagcgtag agaagttgca
     2161 tgtgattgat cggcaaacac aagaggaatt ggagacagtg gagatctcag aagttcagct
     2221 ggtctacaat agcagccatt tcaaatcact agccactgga ggaaatgtta gccaggcact
     2281 ggctttggtt ggagagaagg cttgttatca atccatcagt agctatggtg gtcagatctt
     2341 ttatttgggg acaaaatctg tttatgtgat gatgctgagg agctggagag agagagtgga
     2401 tcatctcctg aaacaagatt gtcttacaga agcgttggct cttgcgtggt ctttccatga
     2461 aggaaaagca aaagcagtag tgggattatc aggggatgcc agtaagcgaa aggctattgt
     2521 tgcagaccgg atggtagaaa tcctattcca ttatgcagat cgagctctga aaaagtgccc
     2581 agaccaagga aaaatccaag tgatggagca gcattttcag gatatggtgc ctgtcatagt
     2641 tgattactgc cttctgctgc agcgaaagga tcttttattt agtcagatgt atgataaatt
     2701 aagtgagaat tcagtggcca aaggagtatt tttggagtgc cttgagccat atattttaag
     2761 tgataaattg gtgggaatca caccccaagt aatgaaagac ttgattgttc atttccaaga
     2821 taaaaaatta atggaaaatg tggaagcgct cattgtacat atggatatca ccagcctaga
     2881 tattcagcag gtagttctca tgtgttggga aaatcgttta tatgatgcta tgatctatgt
     2941 ctacaacaga ggcatgaatg aatttattag tccaatggag aaacttttca gagtcattgc
     3001 tcctcctctg aatgcaggaa aaacactaac agatgaacaa gttgttatgg gcaataagct
     3061 ccttgtatat attagctgtt gtctagcagg tcgtgcctat ccccttggtg acatccctga
     3121 agatctggtt cccttggtta aaaaccaggt ttttgaattt ctaattcgcc tgcattcagc
     3181 agaggcttct cctgaggaag aaatctatcc ttacattcgg actttgctac attttgacac
     3241 aagagaattt ctaaatgtat tggcactgac ttttgaagat tttaaaaatg acaagcaagc
     3301 tgtggaatat caacagcgaa ttgtggatat tttgttgaaa gttatggtgg agaattcaga
     3361 ctttaccccc tcacaagtag gatgtctctt taccttcctt gctcggcagc ttgcaaagcc
     3421 tgacaacacc ttgtttgtaa acagaacact ttttgatcag gtccttgaat tcctttgtag
     3481 tcctgacgat gactcccgac actctgaaag acagcaggtc cttttagaat tgctgcaggc
     3541 tggaggcata gttcaatttg aagagagtcg actcatccgg atggcagaaa aagctgagtt
     3601 ctatcaaatt tgtgaattta tgtatgaaag agaacaccaa tatgataaaa ttattgattg
     3661 ctacttacgt gaccctctgc gagaggaaga agtctttaat tacattcaca atatcttatc
     3721 cattcccgga cacagtgcag aggagaagca gtctgtatgg cagaaagcaa tggatcatat
     3781 tgaggaactc gtgtccctga agccttgtaa agctgcggag ctggttgcca cccacttttc
     3841 tggacatatt gaaacggtca ttaaaaaact tcagaaccag gttttgcttt tcaaattttt
     3901 gaggagtctt cttgacccaa gggaaggtat tcatgtaaat caagaattac tgcaaatatc
     3961 tccttgtatc acagagcagt tcattgagct gttgtgtcag ttcaacccaa cccaagttat
     4021 agagactctg caagtccttg agtgctaccg tctggaagaa actattcaga ttactcagaa
     4081 gtatcaactt catgaagtca ccgcttatct attggaaaag aaaggagata ttcatggtgc
     4141 cttcctaata atgttagaga gactacaaag caaacttcaa gaggtaacac atcaaggtga
     4201 aaataccaaa gaggatccct cattgaagga tgttgaagat actatggtgg agaccattgc
     4261 tctttgccag agaaattcac ataatttgaa ccagcagcaa cgtgaggcac tttggtttcc
     4321 gttattggag gcaatgatgg cccctcagaa gctgtccagt tcagccattc ctcatctaca
     4381 ctctgaagct ctgaagtctt tgaccatgca agttttaaat agcatggcag catttattgc
     4441 ccttccatca atcttgcaaa gaatcttaca ggatccagtt tatggaaaag gaaaacttgg
     4501 agaaatccag ggacttatct tgggaatgtt agataccttt aactatgaac aaaccctgct
     4561 ggaaacaaca accagccttc taaaccaaga tctccattgg tcattgtgta acctgagagc
     4621 ttcggtcacc agaggactga atcccaaaca agattactgc tctatatgtt tgcagcagta
     4681 caagagacgc caagaaatgg ctgatgaaat aattgtcttt agctgtggcc atttgtatca
     4741 ctcattctgc ctacaaaaca aagaatgcac tgtggaattt gagggccaaa caagatggac
     4801 atgctacaaa tgcagttcaa gtaacaaagt aggaaaactc agtgaaaatt catctgaaat
     4861 taaaaaggga aggataaccc catcacaggt aaaaatgtct ccatcgtatc atcagtccaa
     4921 aggggatccc actgctaaaa agggaacctc agaacctgtt ctggatccac agcaaatcca
     4981 agcatttgat cagctttgcc gtctctaccg aggaagctcc aggctggctc tcctcacgga
     5041 actctcccag aatcgcagca gcgagagcta taggccattc agtggctcgc agagtgctcc
     5101 tgctttcaac agcatcttcc agaatgagaa cttccagctg cagctcattc ctccacctgt
     5161 gactgaggat gaattcggtg gaggcggatc aggaggcggt ggatctatgg tgagcaaggg
     5221 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     5281 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     5341 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     5401 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     5461 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     5521 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     5581 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     5641 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     5701 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     5761 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     5821 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     5881 ccactccacc ggcggcatgg acgagctgta caagtaggcg gccgctcgag tctagagggc
     5941 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6001 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6061 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     6121 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     6181 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     6241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6301 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6421 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6481 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6541 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6601 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6661 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6721 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6841 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6901 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     6961 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     7021 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     7081 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     7141 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     7201 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     7261 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     7321 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     7381 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     7441 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     7501 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     7561 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     7621 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7681 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7741 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7801 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     7861 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     7921 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7981 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     8041 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     8101 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8161 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     8221 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     8281 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     8341 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     8401 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     8461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     9001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     9061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     9121 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     9181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     9241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     9301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     9361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     9421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     9481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     9541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9661 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     9721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9961 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    10021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    10081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    10141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    10201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    10261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    10321 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
    10381 gatctcccga tcccctatgg t
//
