LOCUS       40924_4060        7820 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pADH70, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7820)
  AUTHORS   Nobile CJ.
  TITLE     The Transcriptional Circuitry Controlling Candida albicans Biofilm 
            Formation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7820)
  AUTHORS   Hernday AD.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-SEP-2011) Biochemistry and Biophysics, UCSF, Genentech
            Hall N374, 600 16th Street, San Francisco, CA 94143, USA
REFERENCE   3  (bases 1 to 7820)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7820)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-SEP-2011) Biochemistry and Biophysics, UCSF, Genentech Hall 
            N374, 600 16th Street, San Francisco, CA 94143, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(99..204)
                     /label=AmpR promoter
     rep_origin      231..686
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     827..843
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        853..871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             925..954
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             961..990
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1006..1035
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1042..1071
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1078..1107
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1123..1152
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1159..1188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(1221..1254)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             1813..3081
                     /codon_start=1
                     /label=FLP
                     /note="site-specific recombinase"
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             join(3977..3986,4641..5203)
                     /codon_start=1
                     /gene="SAT1"
                     /product="SAT1"
                     /label=SAT1
                     /note="nourseothricin resistance marker"
                     /protein_id="AEX01746.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     gene            3977..5203
                     /gene="SAT1"
                     /label=SAT1
     protein_bind    complement(5341..5374)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        complement(5420..5438)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5459..5475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5483..5499)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5507..5537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5552..5573)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5861..6449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6669..6771
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6772..7428
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
       61 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
      121 tgaagcattt atcaagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      181 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
      241 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
      301 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
      361 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
      421 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
      481 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
      541 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
      601 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
      661 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa
      721 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
      781 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      841 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag
      901 cggatccccg ggttaattaa cggtgaacaa aagctaatct ccgaggaaga cttgaacggt
      961 gaacaaaaat taatctcaga agaagacttg aacggactcg acggtgaaca aaagttgatt
     1021 tctgaagaag atttgaacgg tgaacaaaag ctaatctccg aggaagactt gaacggtgaa
     1081 caaaaattaa tctcagaaga agacttgaac ggactcgacg gtgaacaaaa gttgatttct
     1141 gaagaagatt tgaacggtga acaaaagcta atctccgagg aagacttgaa cggtgaacaa
     1201 aaattaatca atcactcgag gaagttccta tactttctag agaataggaa cttcggatcc
     1261 aataatgatt ggtttgatat ttttgtctag taccatctgt accattacac ttaaattatc
     1321 tttatatctg tctaactcga ctgtctggat ttcattgatg tagtcgtatg catcgttagt
     1381 tccaaaaaat attgtcatca atttgatatt ggtttccgac tctaaaattt ttggaagaat
     1441 ttgtctagcg tgctctgagt tgtagccact gaaaccacgg ttaataacat ccaattttcg
     1501 gatatacaca ttctgtaatg ctggatgaaa gccatactgg gtacaactaa actgggtgat
     1561 ggagtcaccg aacaacacaa atttaccgta ttccatgatt gctatggttg agaatttttt
     1621 ttttttcttg tcccacgcca tttttcaaat tatgcagttg agaatgttag tttttgtgta
     1681 caccccgttc gctgaatatt tcggaataat tcaaagattg gggagtgggg gaggcgatag
     1741 acgaagacac ggtataaaaa tgggcaaaat tttccccaac tttttgcagt ggtttaacta
     1801 ataatcgtcg acatgccaca atttgatata ttatgtaaaa caccacctaa ggtgcttgtt
     1861 cgtcagtttg tggaaaggtt tgaaagacct tcaggtgaga aaatagcatt atgtgctgct
     1921 gaactaacct atttatgttg gatgattaca cataacggaa cagcaatcaa gagagccaca
     1981 ttcatgagct ataatactat cataagcaat tcgttgagtt tcgatattgt caataaatca
     2041 ctccagttta aatacaagac gcaaaaagca acaattttgg aagcctcatt aaagaaattg
     2101 attcctgctt gggaatttac aattattcct tactatggac aaaaacatca atctgatatc
     2161 actgatattg taagtagttt gcaattacag ttcgaatcat cggaagaagc agataaggga
     2221 aatagccaca gtaaaaaaat gcttaaagca cttctaagtg agggtgaaag catctgggag
     2281 atcactgaga aaatactaaa ttcgtttgag tatacttcga gatttacaaa aacaaaaact
     2341 ttataccaat tcctcttcct agctactttc atcaattgtg gaagattcag cgatattaag
     2401 aacgttgatc cgaaatcatt taaattagtc caaaataagt atttgggagt aataatccag
     2461 tgtttagtga cagagacaaa gacaagcgtt agtaggcaca tatacttctt tagcgcaagg
     2521 ggtaggatcg atccacttgt atatttggat gaatttttga ggaattctga accagtccta
     2581 aaacgagtaa ataggaccgg caattcttca agcaataaac aggaatacca attattaaaa
     2641 gataacttag tcagatcgta caataaagct ttgaagaaaa atgcgcctta ttcaatcttt
     2701 gctataaaaa atggcccaaa atctcacatt ggaagacatt tgatgacctc atttctttca
     2761 atgaagggcc taacggagtt gactaatgtt gtgggaaatt ggagcgataa gcgtgcttct
     2821 gccgtggcca ggacaacgta tactcatcag ataacagcaa tacctgatca ctacttcgca
     2881 ctagtttctc ggtactatgc atatgatcca atatcaaagg aaatgatagc attgaaggat
     2941 gagactaatc caattgagga gtggcagcat atagaacagc taaagggtag tgctgaagga
     3001 agcatacgat accccgcatg gaatgggata atatcacagg aggtactaga ctacctttca
     3061 tcctacataa atagacgcat ataagagtga aattctggaa atctggaaat ctggttttgt
     3121 attcttgtta ttcttctttt tgttattaca tatataactt gttacttttt taaaaaaatc
     3181 tttgtatatt ttataaatat ataaaactaa atttaagaaa aagagaaaaa tgttttattt
     3241 gagagattga aattttactt gaatttagct tagcttttat aaagtattat tatgtaaaaa
     3301 aacaaaacaa atatacatta aaaagttaag actataaaat agccacccaa ggcatttcta
     3361 tatcttgttg ttgttgtttt catcttctgt atcagaggaa cttattttat tattttcgtc
     3421 acgggtattt tctcttgttt gatgattcat cccattcatt ccatcataaa atgtcgagcg
     3481 tcaaaactag agaataataa agaaaacgat cttttcaaaa agaaaaaacc ttttagtttt
     3541 cctttgttgt tgttgtgggt gtgtgctatt tatattatat agtttactca taataccata
     3601 aaatattcgg tttgattagg ttattttaat aagctaattt gtttctaatc gtgtaattta
     3661 tgctgtgtat attaagtagt gtgtgcactg cccaaaaatg tttgttgttt atagtcggtt
     3721 aaagagaaaa aagaaaaaaa gatccataca cacacgttaa ttagttgttc aacgtaatac
     3781 actcatattt tgttcttatt tgctttcggt cgctgttctc accaagattt attgccaacg
     3841 aaacaatttt tttttatata ttttcagatt tttctttttt tcctttcctt tccttttcta
     3901 attttcactc ctggttttct ttctttctta gaaacattat ctcgatatta atattaaaaa
     3961 aatataatca ttcaaaatgg acggtggtat gttttagttt agcttcaatt ctaattgatt
     4021 gattaatcag ttgattggtt tcaatatgac aaatgggtag ggtgggaaaa cttcattttc
     4081 aattcagatc aaactttttt gttgtcgaca taatatttct cgtttgggat gttactgtca
     4141 cattaataat acacacacat cagcttataa ttttgaaagt aatttatcag atatgttgtg
     4201 acgatcaatg gaaatggcta acttcaatgt atctgttctt cccctttttc aaagttcacg
     4261 ttttttgatt gattgattga tctgtcggca gtggtttcaa aaccattcgg tgagtaatcc
     4321 tatcaatcaa tgttacgaca aaaggctcaa tattcaaaat tgcaatgttt tatgttttcc
     4381 tacgtgtact tgtgcaaggc aattgattca acattgcttt tggtgtttga cgagtttcta
     4441 gtttggactt gtgttgttat ctgggctata cagatttccc ggctcactat gaattttttt
     4501 tttcgacgct cagtgcacac aactataaac aacacaaaca caaacacagc aagaaaaaaa
     4561 aaaaacgaac attgaattga aaccaagcca actgaaaaat tccttattta aatgactgtc
     4621 atactaaccc atttttatag aagaagttgc tgctttagtt atcgataacg gttctcatat
     4681 gaaaatttcg gtgatccctg agcaggtggc ggaaacattg gatgctgaga accatttcat
     4741 tgttcgtgaa gtgttcgatg tgcacctatc cgaccaaggc tttgaactat ctaccagaag
     4801 tgtgagcccc taccggaagg attacatctc ggatgatgac tctgatgaag actctgcttg
     4861 ctatggcgca ttcatcgacc aagagcttgt cgggaagatt gaactcaact caacatggaa
     4921 cgatctagcc tctatcgaac acattgttgt gtcgcacacg caccgaggca aaggagtcgc
     4981 gcacagtctc atcgaatttg cgaaaaagtg ggcactaagc agacagctcc ttggcatacg
     5041 attagagaca caaacgaaca atgtacctgc ctgcaatttg tacgcaaaat gtggctttac
     5101 tctcggcggc attgacctct tcacgtataa aactagacct caagtctcga acgaaacagc
     5161 gatgtactgg tactggttct cgggagcaca ggatgacgcc taacatatgt gaagtgtgaa
     5221 gggggagatt ttcactttat tagatttgta tatatgtata ataaataaat aaataagtta
     5281 aataaataat tagataaggg tggtaattat tactatttac aatcaaaggt ggtcctgcag
     5341 gaagttccta tactttctag agaataggaa cttcggtgga ggtgcggccg ccaccgcggt
     5401 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5461 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5521 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5581 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5641 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5701 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5761 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5821 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5881 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5941 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6001 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6061 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6121 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6181 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6241 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6301 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6361 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6421 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6481 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6541 tcttcaccta gatccttttc gaccgaataa atacctgtga cggaagatca cttcgcagaa
     6601 taaataaatc ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa
     6661 tgagacgttg atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     6721 ccgggcgtat tttttgagtt gtcgagattt tcaggagcta aggaagctaa aatggagaaa
     6781 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     6841 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     6901 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     6961 gcccgcctga tgaatgctca tccggaatta cgtatggcaa tgaaagacgg tgagctggtg
     7021 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     7081 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     7141 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     7201 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     7261 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     7321 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     7381 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     7441 ggcagttatt ggtgccctta aacgcctggt tgctacgcct gaataagtga taataagcgg
     7501 atgaatggca gaaattcgaa agcaaattcg acccggtcgt cggttcaggg cagggtcgtt
     7561 aaatagccgc ttatgtctat tgctggttta ccggtttatt gactaccgga agcagtgtga
     7621 ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg gaggtaataa
     7681 ttgacgatat gatccttttt ttctgatcaa aagtgctcat cattggaaaa cgttcttcgg
     7741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7801 cacccaactg atcttcagca
//
