LOCUS       V025882                 8849 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025882
VERSION     V025882
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             874..1590
                     /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"
     CDS             1660..4221
                     /label="VPS41(NM_014396)"
                     /note="VPS41(NM_014396)"
                     /gene="VPS41"
     CDS             4228..4254
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4258..4284
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4288..4314
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4416..4640
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4686..5114
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5128..5457
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5524..6318
                     /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    6492..6613
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6662..6678)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6686..6702
                     /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(6710..6740)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6755..6776
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7064..7649)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7820..8680)
                     /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(8681..8785)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tggcggaagc agaggagcag
     1681 gaaactgggt cccttgaaga atctacagat gagtctgagg aagaagagag cgaagaggaa
     1741 cccaagctga agtatgaaag gctttccaat ggggtaactg aaatacttca gaaggatgca
     1801 gctagctgca tgacagtcca tgacaagttt ttggcattgg gcacacatta tggcaaggtt
     1861 tatttacttg atgtccaggg gaacatcact cagaagtttg atgtaagtcc tgtgaagata
     1921 aatcagatta gcttggatga aagtggagag cacatgggtg tgtgttcaga ggatggcaag
     1981 gtgcaggtat ttggactgta ttctggagaa gaatttcacg agacttttga ctgtcccatt
     2041 aaaattattg ctgtgcaccc acatttcgtg agatccagtt gcaagcagtt tgtgaccgga
     2101 gggaagaagc tgctactgtt tgaacggtct tggatgaaca gatggaagtc tgctgttctg
     2161 catgaagggg aagggaacat aaggagtgtg aagtggagag gccatctgat tgcttgggcc
     2221 aataatatgg gtgtgaagat ttttgacatc atctcaaagc aaagaatcac caatgtgccc
     2281 cgggatgata taagtcttcg cccagacatg tatccctgca gcctctgctg gaaggacaat
     2341 gtgacactga ttattggctg ggggacttct gtcaaggtgt gctcagtgaa ggaacggcat
     2401 gccagtgaaa tgagggattt gccaagtcga tatgttgaaa tagtgtctca gtttgaaact
     2461 gaattctaca tcagtggact tgcacctctc tgtgatcagc ttgttgtact ttcgtatgta
     2521 aaggagattt cagaaaaaac ggaaagagaa tactgtgcca ggcctagact ggacatcatc
     2581 cagccacttt ctgagacttg tgaagagatc tcttctgatg ctttgacagt cagaggcttt
     2641 caggagaatg aatgtagaga ttatcattta gaatactctg aaggggaatc acttttttac
     2701 atcgtgagtc cgagagatgt tgtagtggcc aaggaacgag accaagatga tcacattgac
     2761 tggctccttg aaaagaagaa atatgaagaa gcattgatgg cagctgaaat tagccaaaaa
     2821 aatattaaaa gacataagat tctggatatt ggcttggcat atataaatca cctggtggag
     2881 agaggagact atgacatagc agcacgcaaa tgccagaaaa ttcttgggaa aaatgcagca
     2941 ctctgggaat atgaagttta taaatttaaa gaaattggac agcttaaggc tattagtcct
     3001 tatttgccaa gaggtgatcc agttctgaaa ccactcatct atgaaatgat cttacatgaa
     3061 tttttggaga gtgattatga gggttttgcc acattgatcc gagaatggcc tggagatctg
     3121 tataataatt cagtcatagt tcaagcagtt cgggatcatt tgaagaaaga tagtcagaac
     3181 aagactttac ttaaaaccct ggcagaattg tacacctatg acaagaacta tggcaatgct
     3241 ctggaaatat acttaacatt aagacataaa gacgtttttc agttgatcca caagcataat
     3301 cttttcagtt ctatcaagga taaaattgtt ttattaatgg attttgattc agagaaagct
     3361 gttgacatgc ttttggacaa tgaagataaa atttcaatta aaaaggtagt ggaagaattg
     3421 gaagacagac cagagctaca gcatgtgtat ttgcataagc ttttcaagag agaccaccat
     3481 aaggggcagc gttaccatga aaaacagatc agtctttatg ctgaatatga tcgaccaaac
     3541 ttacttccct ttctccgaga cagtacccat tgcccacttg aaaaggctct tgagatctgt
     3601 caacagagaa actttgtaga agagacagtt tatcttctga gccgaatggg taatagccga
     3661 agtgccctga agatgattat ggaggaatta catgatgttg ataaagcaat cgaatttgcc
     3721 aaggagcaag atgatggaga gctgtgggaa gatttgattt tatattccat tgacaaacca
     3781 ccatttatta ctggcttgtt aaacaacatt ggcacacatg ttgacccaat tctactgatt
     3841 caccgtatta aggaaggaat ggagatcccc aatttgagag attccttggt taaaattctg
     3901 caagactaca atttgcaaat tctgcttcgt gaaggctgca agaagattct cgtagctgac
     3961 tctttgtcct tactgaagaa aatgcaccga actcaaatga aaggtgttct tgttgatgag
     4021 gagaacatct gtgagtcgtg cctttcccct attcttccat cagatgcagc taagcccttc
     4081 agcgtggtgg tcttccattg ccggcacatg ttccacaagg agtgcctgcc catgcccagc
     4141 atgaactctg ctgcacagtt ctgcaacatc tgcagtgcta agaaccgtgg accaggaagt
     4201 gcaattttgg agatgaaaaa aggtacctac ccatacgacg tacctgatta tgcaacttat
     4261 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcataggat
     4321 ccactagtcc agtgtggtgg aattctgcag atatccagca cagtggcggc cgctcgagtc
     4381 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4441 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4501 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4561 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4621 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4681 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4741 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4801 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4861 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4921 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4981 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5041 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5101 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5161 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5281 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5341 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5401 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5461 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5521 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5581 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5641 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5701 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5761 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5821 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5881 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5941 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6001 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6061 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6121 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6181 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6241 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6301 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6361 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6421 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6481 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6541 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6601 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6661 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     7621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7741 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7801 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7861 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7921 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7981 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8041 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8101 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8161 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8221 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8281 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8341 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8401 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8461 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8521 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8581 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8641 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8701 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8761 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8821 gatcgggaga tctcccgatc ccctatggt
//
