LOCUS       V043018                 7946 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043018
VERSION     V043018
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..3466
                     /label="Vps41(NM_172120)"
                     /note="Vps41(NM_172120)"
                     /gene="Vps41"
     polyA_signal    3513..3737
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3783..4211
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4225..4554
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4621..5415
                     /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    5589..5710
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5759..5775)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5783..5799
                     /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(5807..5837)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5852..5873
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6161..6746)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6917..7777)
                     /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(7778..7882)
                     /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 gcggaagcag aggagcagga aaccgagtct cttgaagaat ctacggatga
      961 gtcagaggaa gaaagtgaag aggaacccaa actgaagtat gaaaggcttt ccaatggggt
     1021 gacagagatt cttcagaagg atgcagcaag ctgtatgacg gtccatgaca agtttttggc
     1081 actgggaaca cattatggca aggtttattt acttgatgtc cagggaaaca tcactcagaa
     1141 gtttgatgta agtcctgtga agataaacca aattagcttg gatgacagtg gagaacacat
     1201 gggagtgtgt tcagaggatg gcaagctgca ggtctttggg ctgtattctg gagaagaatt
     1261 tcacgagact tttgactgtc ccattaaaat cattgctgtg cacccacagt tcgtgagatc
     1321 cagttgtaag cagtttgtga ctggcgggaa aaagctgctg ctgtttgaac ggacctggat
     1381 gaacagatgg aagtcttctg tgctacacga aggggaagga aacataagga gtgtgaagtg
     1441 gagaggccat ctgattgctt gggccaacaa tatgggtgtg aaggtttttg acatcacctc
     1501 aaagcaaaga atcagcaatg tgccccgaga tgacataagt cttcgcccag acatgtaccc
     1561 ctgcagcctc tgctggaagg acaatgtgac actcattatt ggctggggaa cttccatcaa
     1621 gatatgctca gtgaaggaac gccacgccag tgagatgagg gatctgccaa gccgatatgt
     1681 tgagatagtg tctcagtttg aaactgaatt ctacattagt ggacttgcac ctctctgtga
     1741 tcagcttgtt gtactttcct acgtaaagga ggtttcagaa aaaacggaaa gagaatactg
     1801 tgcaagacct agactggaca tcatccagcc acttccggag acttgtgaag agatctcctc
     1861 cgatgctttg acagtcagag gctttcagga gaatgaatgt agggattacc acttagaata
     1921 ctccgagggg gaatcactct tctatgtcgt gagtccaaga gatgtcgtcg tagccaagga
     1981 acgagaccag gatgatcaca tcgattggct ccttgagaag aagaaatatg aagaagcttt
     2041 gatggctgct gaaattagcc aaaggaacat taaacgacat aagattctgg atattgggtt
     2101 ggcatatgta aaccacctgg tggagagagg agagtacgac atggctgcaa gaaaatgcca
     2161 gaaaattctt gggaaaaatg catccctctg ggaatacgaa gtttacaaat ttaaagaaat
     2221 tgggcagctt aaagcaatta gtccctattt acctcgaggg gacccagttc tgaaaccact
     2281 catctatgaa atgattttgc acgaatttct ggaaagtgac tatgagggct ttgccacatt
     2341 gatcagagag tggcctggag atctatataa caattccgtg atagttcagg cagttcgaga
     2401 ccacctgaag aaggacagtc agaacaaaac gttattgaaa acactggcag aattgtacac
     2461 ctatgacaaa aattatggca atgctctgga aatatactta acgttaagac ataaagatgt
     2521 tttccaactg atacacaagc ataatctttt cagctccatc aaggataaaa ttgtgttatt
     2581 aatggatttt gattcagaga aagctgttga catgcttttg gacaatgaag ataaaatttc
     2641 aattaaaaaa gtggtagaag aattagaaga cagaccagaa ctgcagcatg tgtatttgca
     2701 caagctcttc aagagagacc atcataaggg acagcgctac catgaaaagc agatcagtct
     2761 gtatgccgag tatgaccgac caaacctgct tccctttctc cgagacagta cccattgccc
     2821 acttgaaaag gcacttgaga tctgtcaaca aagaaacttt gtagaagaga cagtttacct
     2881 tctgagccga atgggaaaca gtagaagtgc cctcaagatg attatggagg aattgcatga
     2941 tgttgataaa gcaattgagt ttgccaagga acaagatgat ggagagctct gggaagatct
     3001 gattctatac tccatcgaca aaccaccatt tatcactggc ttgttgaaca acattggcac
     3061 acatgtggac ccgattctac ttatccaccg aattaaggaa ggaatggaga ttcccaattt
     3121 gagagattcc ttggttaaaa ttctgcaaga ttataacttg cagattctgc ttcgagaagg
     3181 ctgcaagaaa atcctcgtag ctgactcatt gtcgttgctg aaaaaaatgc accgaaccca
     3241 aatgaaaggc gttctggttg atgaagagaa tatctgtgaa tcatgccttt ctcccatcct
     3301 cccaacagat gcagcgaagc ccttcagcgt ggtggtcttc cactgccggc acatgttcca
     3361 caaggagtgc ctgcctatgc cgagcatgaa tgctcctgca caatactgca acatctgtag
     3421 tgctaagaac cgtgggcctg gaagtgccat cctggagatg aagaagtagc tcgagtctag
     3481 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3541 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3601 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3661 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3721 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3781 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3841 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3901 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3961 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4021 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4081 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4141 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4201 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4261 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4321 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4381 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4441 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4501 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4561 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4621 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4681 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4741 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4801 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4861 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4921 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4981 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5041 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5101 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5161 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5221 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5281 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5341 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5401 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5461 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5521 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5581 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5641 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5701 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5761 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5821 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5881 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5941 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6061 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6601 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6661 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7921 cgggagatct cccgatcccc tatggt
//
