LOCUS       V036595                 7640 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036595
VERSION     V036595
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             908..3076
                     /label="VPS52(NM_022553)"
                     /note="VPS52(NM_022553)"
                     /gene="VPS52"
     CDS             3083..3172
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3207..3431
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3477..3905
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3919..4248
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4315..5109
                     /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    5283..5404
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5453..5469)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5477..5493
                     /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(5501..5531)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5546..5567
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5855..6440)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6611..7471)
                     /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(7472..7576)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gccgccgctg cgaccatggc ggctgcggcc cgggaactgg tgttgcgggc
      961 tgggacctca gatatggagg aggaagaggg cccgctggcg ggtggtcctg ggctccagga
     1021 accactgcaa cttggggagt tggatatcac ttctgatgaa ttcatcctgg atgaagtgga
     1081 tgttcacatt caggcaaatc tggaggatga gttagtaaag gaagctctta aaacgggtgt
     1141 agatctccgt cactattcaa agcaagttga gctggagcta cagcagattg aacagaaatc
     1201 cattcgggat tatattcaag agagtgagaa tatagcatct ctacacaacc agatcacagc
     1261 ctgtgatgct gtcctggagc gaatggagca gatgttggga gcttttcaga gtgacctcag
     1321 ctccatcagc tctgagatcc ggacactgca ggaacagtca ggagccatga acattcgact
     1381 tcgaaatcgc caggcagttc gggggaaact tggggagctt gttgatggtc tggtggtgcc
     1441 ttctgctctg gtcacggcaa ttctggaggc tccagtgaca gagcccaggt tcttggagca
     1501 gctacaggag ctggatgcca aggcagccgc agtcagagag caggaagcta gaggcacagc
     1561 agcctgcgca gatgtcagag gcgtgctcga tcggctccgg gtcaaggcag tgacgaagat
     1621 ccgagagttt atcctccaga agatttattc cttcaggaaa cccatgacca actatcagat
     1681 cccccagacg gccctgctga agtacaggtt cttctatcag tttctgctgg gcaatgaacg
     1741 agcaacagca aaggagatca gggatgaata tgtggagacg ctgagcaaga tttacctgtc
     1801 ttactaccgc tcttacctgg ggcggctcat gaaggtgcag tatgaggaag tcgctgagaa
     1861 agatgatcta atgggtgtgg aagatacagc aaagaaagga ttcttctcaa agccatcgct
     1921 ccgcagcagg aacaccattt tcaccctagg aacccgcggc tctgtcatct cccccactga
     1981 acttgaggcc cccatcctgg tgcctcacac agcgcagcgc ggagagcaga ggtatccatt
     2041 tgaggccctc ttccgcagcc agcactacgc cctcctagac aattcctgcc gcgaatacct
     2101 tttcatctgt gaattttttg ttgtgtctgg cccagctgca cacgacctgt tccatgctgt
     2161 catgggccgt acactcagca tgaccctgaa acacctggat tcttatctag ctgactgcta
     2221 cgatgccatt gctgtttttc tctgtatcca cattgttctc cggttccgta acattgcagc
     2281 aaagagggat gttcctgccc tggacaggta ctgggaacag gtgcttgcct tgctatggcc
     2341 acggtttgaa ctgatcctgg agatgaatgt tcagagcgtc cgaagcactg acccccagcg
     2401 cctagggggg ttggatactc ggccccacta tatcacacgc cgctatgcag agttctcctc
     2461 cgctcttgtc agtatcaacc agacaattcc taatgaacgg accatgcaat tgctgggaca
     2521 gctgcaggtg gaggtggaga attttgtcct ccgagtggca gctgagttct cctcaaggaa
     2581 ggagcagctt gtgtttctga tcaacaacta tgacatgatg ctgggtgtgc tgatggagcg
     2641 ggctgcagat gacagcaaag aggttgagag cttccagcag ctgctcaatg ctcggacaca
     2701 ggaattcatt gaagagttgc tgtctccccc ttttgggggt ttagtggcat ttgtgaagga
     2761 ggctgaggct ttgattgagc gtggacaggc tgagcgactt cgaggggaag aagcccgggt
     2821 aactcagctg atccgtggct ttggtagttc ctggaaatca tcagtggaat ctctgagtca
     2881 ggatgtaatg cggagtttca ccaacttcag aaatggcacc agtatcattc agggagcgct
     2941 gacccagctg atccagctct atcatcgctt ccaccgggtg ctgtcccagc cgcagctccg
     3001 agccctccct gcccgggctg agctcatcaa cattcaccac cttatggtgg agctcaagaa
     3061 gcataagccc aacttcctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa
     3121 actaatcagt gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatgagggcc
     3181 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3241 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3301 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3361 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3421 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3481 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3541 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3601 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3661 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3721 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3781 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3841 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3901 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4081 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4141 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4201 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4261 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4321 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4381 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4441 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4501 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4561 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4621 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4681 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4741 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4801 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4861 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4921 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4981 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5041 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5101 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5161 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5221 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5281 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5341 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5401 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5461 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5521 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5581 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5641 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5761 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6361 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6421 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6481 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6541 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6661 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6721 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6781 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6841 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6901 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6961 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7021 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7081 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7141 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7201 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7261 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7321 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7381 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7441 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7561 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7621 atctcccgat cccctatggt
//
