LOCUS       V022799                 9715 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022799
VERSION     V022799
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..4395
                     /label="CAND1(NM_018448)"
                     /note="CAND1(NM_018448)"
                     /gene="CAND1"
     CDS             4402..4467
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4491..5079
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5145..5193
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5395..5823
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5837..6166
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6233..7027
                     /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    7203..7324
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7373..7389)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7397..7413
                     /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(7421..7451)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7466..7487
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7775..8363)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8534..9394)
                     /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(9395..9499)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gagcgcctcg
      721 taccacattt ccaatttgct ggaaaaaatg acatccagcg acaaggactt taggtttatg
      781 gctacaaatg atttgatgac ggaactgcag aaagattcca tcaagttgga tgatgatagt
      841 gaaaggaaag tagtgaaaat gattttgaag ttattggaag ataaaaatgg agaggtacag
      901 aatttagctg tcaaatgtct tggtccttta gtgagtaaag tgaaagaata ccaagtagag
      961 acaattgtag ataccctctg cactaacatg ctttctgata aagaacaact tcgagacatt
     1021 tcaagtattg gtcttaaaac agtaattgga gaacttcctc cagcttccag tggctctgca
     1081 ttagctgcta atgtatgtaa aaagattact ggacgtctta caagtgcaat agcaaaacag
     1141 gaagatgtct ctgttcagct agaagccttg gatattatgg ctgatatgtt gagcaggcaa
     1201 ggaggacttc ttgttaattt ccatccttca attctgacct gtctacttcc ccagttgacc
     1261 agccctagac ttgcagtgag gaaaagaacc attatcgctc ttggccatct ggttatgagc
     1321 tgtggaaata tagtttttgt agatcttatt gaacatctgt tgtcagagtt gtccaaaaat
     1381 gattctatgt caacaacaag aacctacata caatgtattg ctgctattag taggcaagct
     1441 ggtcatagaa taggtgaata ccttgagaag ataattcctt tggtggtaaa attttgcaat
     1501 gtagatgatg atgaattaag agagtactgt attcaagcct ttgaatcatt tgtaagaaga
     1561 tgtcctaagg aagtatatcc tcatgtttct accattataa atatttgtct taaatatctt
     1621 acctatgatc caaattataa ttacgatgat gaagatgaag atgaaaatgc aatggatgct
     1681 gatggtggtg atgatgatga tcaagggagt gatgatgaat acagtgatga tgatgacatg
     1741 agttggaaag tgagacgtgc agctgcgaag tgcttggatg ctgtagttag cacaaggcat
     1801 gaaatgcttc cagaattcta caagaccgtc tctcctgcac taatatccag atttaaagag
     1861 cgtgaagaga atgtaaaggc agatgttttt cacgcatacc tttctctttt gaagcaaact
     1921 cgtcctgtac aaagttggct atgtgaccct gatgcaatgg agcagggaga aacaccttta
     1981 acaatgcttc agagtcaggt tcccaacatt gttaaagctc ttcacaaaca gatgaaagaa
     2041 aaaagtgtga agacccgaca gtgttgtttt aacatgttaa ctgagctggt aaatgtatta
     2101 cctggggccc taactcaaca cattcctgta cttgtaccag gaatcatttt ctcactgaat
     2161 gataaatcaa gctcatcgaa tttgaagatc gatgctttgt catgtctata cgtaatcctc
     2221 tgtaaccatt ctcctcaagt cttccatcct cacgttcagg ctttggttcc tccagtggtg
     2281 gcttgtgttg gagacccatt ttacaaaatt acatctgaag cacttcttgt tactcaacag
     2341 cttgtcaaag taattcgtcc tttagatcag ccttcctcgt ttgatgcaac tccttatatc
     2401 aaagatctat ttacctgtac cattaagaga ttaaaagcag ctgacattga tcaggaagtc
     2461 aaggaaaggg ctatttcctg tatgggacaa attatttgca accttggaga caatttgggt
     2521 tctgacttgc ctaatacact tcagattttc ttggagagac taaagaatga aattaccagg
     2581 ttaactacag taaaggcatt gacactgatt gctgggtcac ctttgaagat agatttgagg
     2641 cctgttctgg gagaaggggt tcctatcctt gcttcatttc ttagaaaaaa ccagagagct
     2701 ttgaaactgg gtactctttc tgcccttgat attctaataa aaaactatag tgacagcttg
     2761 acagctgcca tgattgatgc agttctagat gagctcccac ctcttatcag cgaaagtgat
     2821 atgcatgttt cacaaatggc catcagtttt cttaccactt tggcaaaagt atatccctcc
     2881 tccctttcaa agataagtgg atccattctc aatgaactta ttggacttgt gagatcaccc
     2941 ttattgcagg ggggagctct tagtgccatg ctagactttt tccaagctct ggttgtcact
     3001 ggaacaaata atttaggata catggatttg ttgcgcatgc tgactggtcc agtttactct
     3061 cagagcacag ctcttactca taagcagtct tattattcca ttgccaaatg tgtagctgcc
     3121 cttactcgag catgccctaa agagggacca gctgtagtag gtcagtttat tcaagatgtc
     3181 aagaactcaa ggtctacaga ttccattcgt ctcttagctc tactttctct tggagaagtt
     3241 gggcatcata ttgacttaag tggacagttg gaactaaaat ctgtaatact agaagctttc
     3301 tcatctccta gtgaagaagt caaatcagct gcatcctatg cattaggcag cattagtgtg
     3361 ggcaaccttc ctgaatatct gccgtttgtc ctgcaagaaa taactagtca acccaaaagg
     3421 cagtatcttt tacttcattc cttgaaggaa attattagct ctgcatcagt ggtgggcctt
     3481 aaaccatatg ttgaaaacat ctgggcctta ttactaaagc actgtgagtg tgcagaggaa
     3541 ggaaccagaa atgttgttgc tgaatgtcta ggaaaactca ctctaattga tccagaaact
     3601 ctccttccac ggcttaaggg gtacttgata tcaggctcat catatgcccg aagctcagtg
     3661 gttacggctg tgaaatttac aatttctgac catccacaac ctattgatcc actgttaaag
     3721 aactgcatag gtgatttcct aaaaactttg gaagacccag atttgaatgt gagaagagta
     3781 gccttggtca catttaattc agcagcacat aacaagccat cattaataag ggatctattg
     3841 gatactgttc ttccacatct ttacaatgaa acaaaagtta gaaaggagct tataagagag
     3901 gtagaaatgg gtccatttaa acatacggtt gatgatggtc tggatattag aaaggcagca
     3961 tttgagtgta tgtacacact tctagacagt tgtcttgata gacttgatat ctttgaattt
     4021 ctaaatcatg ttgaagatgg tttgaaggac cattatgata ttaagatgct gacattttta
     4081 atgttggtga gactgtctac cctttgtcca agtgcagtac tgcagaggtt ggaccgactt
     4141 gttgagccat tacgtgcaac atgtacaact aaggtaaagg caaactcagt aaagcaggag
     4201 tttgaaaaac aagatgaatt aaagcgatct gccatgagag cagtagcagc actgctaacc
     4261 attccagaag cagagaagag tccactgatg agtgaattcc agtcacagat cagttctaac
     4321 cctgagctgg cggctatctt tgaaagtatc cagaaagatt catcatctac taacttggaa
     4381 tcaatggaca ctagtctcga ggactacaaa gaccatgacg gtgattataa agatcatgac
     4441 atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc
     4501 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4561 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4621 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4681 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4741 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4801 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4861 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4921 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4981 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     5041 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac
     5101 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5161 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5221 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5281 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5341 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     5401 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5461 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5521 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     5581 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5641 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5701 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5761 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5821 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5881 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5941 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6001 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6061 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     6121 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     6181 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     6241 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6301 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6361 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     6421 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6481 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6541 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6601 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6661 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6721 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6781 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6841 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6901 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6961 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7021 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     7081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     7141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     7201 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7261 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7321 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7381 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7441 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7501 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7561 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     8221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9541 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     9601 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     9661 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
