LOCUS       V035438                 9736 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035438
VERSION     V035438
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..4428
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4432..4458
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4462..4488
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4512..5100
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5166..5214
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5416..5844
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5858..6187
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6254..7048
                     /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    7224..7345
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7394..7410)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7418..7434
                     /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(7442..7472)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7487..7508
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7796..8384)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8555..9415)
                     /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(9416..9520)
                     /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 gtacccatac gacgtacctg attatgcaac ttatccttat
     4441 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata gtgagggccc
     4501 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4561 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4621 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4681 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4741 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4801 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4861 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     4921 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4981 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5041 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5101 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5161 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     5221 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5281 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5341 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     5401 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5461 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5521 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     5581 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5641 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5701 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5761 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     5821 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     5881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5941 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6001 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6061 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6121 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6181 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     6241 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6301 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6361 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6421 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6481 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6541 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6601 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6661 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6721 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6781 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6841 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6901 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6961 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7021 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     7081 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     7141 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     7201 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     7261 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7321 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     7381 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7441 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7501 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7561 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7621 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7681 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7741 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7801 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7861 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7921 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7981 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8041 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8101 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8161 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8221 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8281 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8341 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8401 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8461 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8521 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8581 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8641 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8701 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8761 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8821 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8881 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8941 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9001 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9061 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9121 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9181 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9241 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9301 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9361 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9421 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9481 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9541 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     9601 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     9661 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     9721 gaagaatctg cttagg
//
