LOCUS       Exported                6341 bp ds-DNA     circular SYN 28-MAY-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBKWH_Lam
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6341)
  AUTHORS   Mireia
  TITLE     Direct Submission
  JOURNAL   Exported 23 Sep 2019 from SnapGene 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6341
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 promoter"
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /note="GAL4 DNA binding domain"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1245..1247
                     /codon_start=1
                     /product="start codon for in vitro 
                     transcription/translation"
                     /note="ATG"
                     /translation="M"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     misc_feature    1281..1309
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1310
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /note="GAL4 activation domain"
                     /translation=""
     protein_bind    1317..1341
                     /gene="mutant version of attB"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1344..1838
                     /codon_start=1
                     /gene="LMNA"
                     /gene_synonym="CDCD1; CDDC; CMD1A; CMT2B1; EMD2; FPL; FPLD;
                     FPLD2; HGPS; IDC; LDP1; LFP; LGMD1B; LMN1; LMNC; LMNL1; 
                     PRO1"
                     /product="lamin isoform C"
                     /note="LMNA"
                     /note="isoform C is encoded by transcript variant 2; lamin 
                     A/C-like 1; 70 kDa lamin; prelamin-A/C; renal carcinoma 
                     antigen NY-REN-32"
                     /db_xref="GI:5031875"
                     /db_xref="CCDS:CCDS1131.1"
                     /db_xref="GeneID:4000"
                     /db_xref="HGNC:HGNC:6636"
                     /db_xref="HPRD:01035"
                     /db_xref="MIM:150330"
                     /protein_id="NP_005563.1"
                     /translation="SEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVR
                     EEFKELKARNTKKEGDLIAAQARLKDLEALLNSKEAALSTALSEKRTLEGELHDLRGQV
                     AKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVE
                     ID"
     protein_bind    complement(1840..1864)
                     /gene="mutant version of attB"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1875..1898
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      1903..1950
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1977..2071
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 terminator"
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     misc_feature    2072..2131
                     /note="H1_overlap_site"
     misc_feature    2128..2178
                     /note="Self_overlap_site"
     misc_feature    2179..2200
                     /note="Primer"
     misc_feature    2313..2339
                     /note="primer"
     rep_origin      complement(2493..3081)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3410..3457
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     CDS             complement(3689..4483)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4484..4588)
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    4664..4714
                     /note="Self_overlap_site"
     misc_feature    4711..4770
                     /note="H2_overlap_site"
     promoter        4786..5067
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     CDS             5068..5742
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(5845..6300)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcccgggg atctccacaa
     1321 gtttgtacaa aaaagcaggc ttatctgaag aggtggtcag ccgcgaggtg tccggcatca
     1381 aggccgccta cgaggccgag ctcggggatg cccgcaagac ccttgactca gtagccaagg
     1441 agcgcgcccg cctgcagctg gagctgagca aagtgcgtga ggagtttaag gagctgaaag
     1501 cgcgcaatac caagaaggag ggtgacctga tagctgctca ggctcggctg aaggacctgg
     1561 aggctctgct gaactccaag gaggccgcac tgagcactgc tctcagtgag aagcgcacgc
     1621 tggagggcga gctgcatgat ctgcggggcc aggtggccaa gcttgaggca gccctaggtg
     1681 aggccaagaa gcaacttcag gatgagatgc tgcggcgggt ggatgctgag aacaggctgc
     1741 agaccatgaa ggaggaactg gacttccaga agaacatcta cagtgaggag ctgcgtgaga
     1801 ccaagcgccg tcatgagacc cgactggtgg agattgacaa cccagctttc ttgtacaaag
     1861 tggtaagtaa ctaagatccg tcgacctgca gcggccgcat aactagcata accccttggg
     1921 gcctctaaac gggtcttgag gggttttttg cgcgcttgca gccaagctaa ttccgggcga
     1981 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     2041 aattttaaag tgactcttag gttttaaaac gttatcccta gtttaaacac ccataatacc
     2101 cataatagct gtttgccaat taccctgtta tccctattac ttctaaatct gaggaggaaa
     2161 atcgaatcgc taccttagcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac
     2221 aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt
     2281 tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagg
     2341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     3241 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     3301 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     3361 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     3421 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     3481 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     3541 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     3601 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     3661 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     3721 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3781 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     3841 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3901 tcggcaagca ggcatcgcca tgagtcacga cgagatcctc gccgtcgggc atgctcgcct
     3961 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4021 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4081 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4141 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4201 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4261 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4321 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4381 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4441 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catactcttc ctttttcaat
     4501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4561 agaaaaataa acaaataggg gttccgcgca catctaagaa accattatta tcatgacatt
     4621 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacttatccc tattacttct
     4681 aaatctgagg aggaaaatcg aatcgctacc ttaggaccgt tatagttact atcaccatat
     4741 atgtaactgg tgattctttc gctaccttag agtgcaccat agatcaacga cattactata
     4801 tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta
     4861 tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg
     4921 tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa
     4981 agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg
     5041 attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt
     5101 ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc
     5161 gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt
     5221 attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa
     5281 tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat
     5341 tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc
     5401 ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt
     5461 gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca
     5521 ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa
     5581 agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga
     5641 ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac
     5701 tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat
     5761 ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc
     5821 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     5881 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5941 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6001 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6061 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6121 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6181 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6241 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga t
//
