LOCUS       V015837                 5167 bp    DNA     circular SYN 22-MAY-2021
DEFINITION  Exported.
ACCESSION   V015837
VERSION     V015837
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5167)
  AUTHORS   Foster AD, Chung C-W., Hann MM, Simpson GL, Tavassoli A.
  TITLE     Development of a fluorescent three-hybrid system for the
            identification of protein-protein associators
  JOURNAL   Pept Sci (Hoboken) 113 (1), e24178 (2021)
REFERENCE   2  (bases 1 to 5167)
  AUTHORS   Foster AD.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAY-2020) Chemistry, University of Southampton,
            Building 27, University Road, Southampton SO17 1BJ, United Kingdom
REFERENCE   3  (bases 1 to 5167)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Pept Sci
            (Hoboken)"; date: "2021"; volume: "113"; issue: "1"; pages: "e24178"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (03-MAY-2020) Chemistry, University of Southampton, Building 27,
            University Road, Southampton SO17 1BJ, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..5167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..79
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             80..1159
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    1175..1196
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1405..1433
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1441..1457
                     /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)."
     CDS             1839..2159
                     /label="FKBP"
                     /note="human FK506-binding protein FKBP12"
     CDS             2187..2471
                     /gene="CI"
                     /label="Repressor protein CI"
                     /note="Repressor protein CI from Enterobacteria phage 434.
                     Accession#: P16117"
     CDS             2508..2783
                     /label="FRB"
                     /note="FKBP-rapamycin binding domain of human FRAP"
     terminator      2844..3090
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    3125..3351
                     /label="HK022 attP"
                     /note="attachment site of phage HK022"
     rep_origin      complement(3385..3773)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3878..4735)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4736..4840)
                     /label="AmpR promoter"
     terminator      4930..5016
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5108..5135
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
       61 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      121 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      181 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
      241 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
      301 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
      361 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
      421 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
      481 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
      541 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
      601 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
      661 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
      721 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
      781 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
      841 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
      901 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
      961 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     1021 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     1081 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1141 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     1201 cacaattctc atgtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt
     1261 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt
     1321 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     1381 tggcaaatat tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa
     1441 ttgtgagcgg ataacaattt cacacaggaa acagccagtc cgtttaggtg ttttcacgag
     1501 cacttcacca acaaggacca tagcatatga gtatttcttc cagggtaaaa agcaaaagaa
     1561 ttcagcttgg acttaaccag gctgaacttg ctcaaaaggt ggggacttcc aatgtttcga
     1621 tctcgcagct cgaaagaggt aaaactaagc ggccgcgctt tttaccagaa cttgcgtcag
     1681 ctcttggcgt aagtgttgac tggctgctca atggcacctc tgattcgaat gttagatttg
     1741 ttgggcacgt tgagcccaaa gggaaatatc cgttgattag catggttaga gccggctcgt
     1801 ggtgtgaagc ttgtgaaccc tacgatatca agctcgaggg tgttcaggtt gaaaccatct
     1861 ctccgggtga cggtcgtacc ttcccgaaac gtggtcagac ctgcgttgtt cactacaccg
     1921 gtatgctgga agacggtaaa aaattcgact cttctcgtga ccgtaacaaa ccgttcaaat
     1981 tcatgctggg taaacaggaa gttatccgtg gttgggaaga aggtgttgct cagatgtctg
     2041 ttggtcagcg tgctaaactg accatctctc cggactacgc ttacggtgct accggtcacc
     2101 cgggtatcat cccgccgcac gctaccctgg ttttcgacgt tgaactgctg aaactggaat
     2161 aaggtaccta agaaggagta tacaacatgt ctatttcttc cagggtaaaa agcaaaagaa
     2221 ttcagcttgg acttaaccag gctgaacttg ctcaaaaggt ggggactacc cagcagtcga
     2281 tcgagcagct cgaaaacggt aaaactaagc ggccgcgctt tttaccagaa cttgcgtcag
     2341 ctcttggcgt aagtgttgac tggctgctca atggcacctc tgattcgaat gttagatttg
     2401 ttgggcacgt tgagcccaaa gggaaatatc cattgattag catggttaga gccggctcgt
     2461 ggtgtgaagc ttgtgaaccc tacgatatca aggtcgaccg tgttgctatc ctgtggcacg
     2521 aaatgtggca cgaaggtctg gaagaagctt ctcgtctgta cttcggtgaa cgtaacgtta
     2581 aaggtatgtt cgaagttctg gaaccgctgc acgctatgat ggaacgtggt ccgcagaccc
     2641 tgaaagaaac ctctttcaac caggcttacg gtcgtgacct gatggaagct caggaatggt
     2701 gccgtaaata catgaaatct ggtaacgtta aagacctgac ccaggcttgg gacctgtact
     2761 accacgtttt ccgtcgtatc tcttaagagc tcgaattctc atgtttgaca gcttatcact
     2821 gatcagtgaa ttaatggcga tgacgcatcc tcacgataat atccgggtag gcgcaatcac
     2881 tttcgtctct actccgttac aaagcgaggc tgggtatttc ccggcctttc tgttatccga
     2941 aatccactga aagcacagcg gctggctgag gagataaata ataaacgagg ggctgtatgc
     3001 acaaagcatc ttctgttgag ttaagaacga gtatcgagat ggcacatagc cttgctcaaa
     3061 ttggaatcag gtttgtgcca ataccagtag aaacagacga agaagctagc taatgctctg
     3121 tctcaggtca ctaatactat ctaagtagtt gattcatagt gactggatat gttgcgtttt
     3181 gtcgcattat gtagtctatc atttaaccac agattagtgt aatgcgatga tttttaagtg
     3241 attaatgtta ttttgtcatc ctttaggtga ataagttgta tatttaaaat ctctttaatt
     3301 atcagtaaat taatgtaagt aggtcattat tagtcaaaat aaaatcattt gtcgatttca
     3361 attttgtccc atggctaatt cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt
     3421 gctttgagag gctctaaggg cttctcagtg cgttacatcc ctggcttgtt gtccacaacc
     3481 gttaaacctt aaaagcttta aaagccttat atattctttt ttttcttata aaacttaaaa
     3541 ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac atgagagctt
     3601 agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg ttagccatga
     3661 gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt agtacgtgaa
     3721 acatgagagc ttagtacgta ctatcaacag gttgaactgc tgatcttcag atcctctacg
     3781 ccggacgcat cgtggccgga tcttgcggcc gcaaaaatta aaaatgaagt tttaaatcaa
     3841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3901 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3961 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4021 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4081 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4141 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4201 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4261 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4321 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4381 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4441 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4501 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4561 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4621 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4681 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4741 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4801 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4861 cacctgcatc gatggccccc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     4921 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     4981 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5041 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     5101 attaagcaga aggccatcct gacggatggc ctttttgcgt ggccaggacc caacgctgcc
     5161 cgaaatt
//
