LOCUS       62056_2940        6842 bp DNA     circular SYN 22-MAY-2021
DEFINITION  Integration vector pAH63-AF_THS, complete sequence.
ACCESSION   MT424825
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6842)
  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 6842)
  AUTHORS   Foster AD.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2020) Chemistry, University of Southampton, 
            Building 27, University Road, Southampton SO17 1BJ, United Kingdom
REFERENCE   3  (bases 1 to 6842)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Pept Sci 
            (Hoboken)"; date: "2021"; volume: "113"; issue: "1"; pages: "e24178"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-MAY-2020) Chemistry, University of Southampton, Building 27, 
            University Road, Southampton SO17 1BJ, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..6842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      561..623
                     /label=P22/434
                     /note="P22/434"
                     /regulatory_class="promoter"
     CDS             624..1244
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             1245..1277
                     /label=ssrA tag
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     regulatory      1287..1330
                     /regulatory_class="terminator"
     promoter        1440..1513
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     CDS             1543..2970
                     /label=tdTomato
                     /note="tandem dimeric (pseudo-monomeric) derivative of
                     DsRed (Shaner et al., 2004)"
     CDS             2992..3651
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     CDS             3657..3668
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             3734..4522
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     terminator      4583..4829
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(4953..5186)
                     /label=lambda attP
                     /note="integrase from phage lambda"
     rep_origin      complement(5253..5641)
                     /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(5731..6522)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      6736..6822
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 cgttggtact ggctctaccg taaaatggcc gatcggtctg ggcggtaaag gtaacgacgg
       61 tatcgccgcg ttcgttcagc gtctgccggg tgcaattggt tatgttgaat atgcttacgc
      121 gaagcagaac aacctggcgt acaccaaact gatctccgct gatggtaaac cggttagtcc
      181 gaccgaagaa aacttcgcta atgcagcaaa aggtgcagac tggagcaaaa ccttcgctca
      241 ggatctgacc aaccagaaag gcgaagatgc atggcctatt acctctacca cgttcattct
      301 gatccacaaa gatcagaaga aaccagaaca agacagaagt gctgaaattc ttcgactggg
      361 cgtacaaaac cggggctaaa caggcgaacg acctggatta cgccagcctg ccggatagtg
      421 tagttgaaca ggttcgcgct gtggaagacc aatattaaag acagtagcgg taagccgctg
      481 tactaataaa actccaggcc gggtacggtg ttttacgccg catccggcat tacaaaatga
      541 ctttgtaaac gcgtggatcc atttgacaaa caagatatct taaatgaaaa tacaagaaat
      601 cttaaatatc tagaggagtc gacatgtcca gattagataa aagtaaagtg attaacagcg
      661 cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga
      721 agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg
      781 acgccttagc cattgagatg ttagataggc accatactca cttttgccct ttagaagggg
      841 aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc
      901 atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc
      961 tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg
     1021 cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag
     1081 tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag
     1141 ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga
     1201 tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtctgcagca aacgacgaaa
     1261 actacgcttt agcagcttaa gcatgcataa aacgaaaggc tcagtcgaaa gactgggcct
     1321 ttcgttttat actagtatca gaacgcagaa gcggtctgga cgtctaagaa accattatta
     1381 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgagt
     1441 ccctatcagt gatagagatt gacatcccta tcagtgatag agatactgag cacatcagca
     1501 ggacgcactg accgaattca ttaaagagga gaaaggtacc gcatggtgag caagggcgag
     1561 gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac
     1621 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     1681 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     1741 atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg
     1801 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg
     1861 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc
     1921 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     1981 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg
     2041 aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag
     2101 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
     2161 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg
     2221 gggcatggca ccggcagcac cggcagcggc agctccggca ccgcctcctc cgaggacaac
     2281 aacatggccg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac
     2341 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2401 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     2461 cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag
     2521 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt
     2581 ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag
     2641 atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg
     2701 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag
     2761 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc
     2821 aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc
     2881 cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg
     2941 ttcctgtacg gcatggacga gctgtacaag taggtcgacg gagggtagtg tatgacagag
     3001 cagaaagccc tagtaaagcg tattacaaat gaaaccaaga ttcagattgc gatctcttta
     3061 aagggtggtc ccctagcgat agagcactcg atcttcccag aaaaagaggc agaagcagta
     3121 gcagaacagg ccacacaatc gcaagtgatt aacgtccaca caggtatagg gtttctggac
     3181 catatgatac atgctctggc caagcattcc ggctggtcgc taatcgttga gtgcattggt
     3241 gacttacaca tagacgacca tcacaccact gaagactgcg ggattgctct cggtcaagct
     3301 tttaaagagg ccctaggggc cgtgcgtgga gtaaaaaggt ttggatcagg atttgcgcct
     3361 ttggatgagg cactttccag agcggtggta gatctttcga acaggccgta cgcagttgtc
     3421 gaacttggtt tgcaaaggga gaaagtagga gatctctctt gcgagatgat cccgcatttt
     3481 cttgaaagct ttgcagaggc tagcagaatt accctccacg ttgattgtct gcgaggcaag
     3541 aatgatcatc accgtagtga gagtgcgttc aaggctcttg cggttgccat aagagaagcc
     3601 acctcgccca atggtaccaa cgatgttccc tccaccaaag gtgttcttat gtagcaattg
     3661 agggtaggtt atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     3721 aaagttaaac atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     3781 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     3841 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     3901 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     3961 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     4021 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     4081 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     4141 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     4201 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     4261 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     4321 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     4381 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     4441 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     4501 gatcaccaag gtagtcggca aataagagct cgaattctca tgtttgacag cttatcactg
     4561 atcagtgaat taatggcgat gacgcatcct cacgataata tccgggtagg cgcaatcact
     4621 ttcgtctcta ctccgttaca aagcgaggct gggtatttcc cggcctttct gttatccgaa
     4681 atccactgaa agcacagcgg ctggctgagg agataaataa taaacgaggg gctgtatgca
     4741 caaagcatct tctgttgagt taagaacgag tatcgagatg gcacatagcc ttgctcaaat
     4801 tggaatcagg tttgtgccaa taccagtaga aacagacgaa gaagctagct ttgcactgga
     4861 ttgcgaggct ttgtgcttct ctggagtgcg acaggtttga tgacaaaaaa ttagcgcaag
     4921 aagacaaaaa tcaccttgcg ctaatgctct gttacaggtc actaatacca tctaagtagt
     4981 tgattcatag tgactgcata tatgttgtgt tttacagtat tatgtagtct gttttttatg
     5041 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttttt
     5101 atactaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     5161 tatcagtcaa aataaaatca ttatttgatt tcaattttgt cccactccct gcctctgtca
     5221 tcacgatact gtgatgccat ggctaattcc catgtcagcc gttaagtgtt cctgtgtcac
     5281 tcaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct ggcttgttgt
     5341 ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt ttcttataaa
     5401 acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt gttcaaacat
     5461 gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc ttagtacgtt
     5521 agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat gagagcttag
     5581 tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgctg atcttcagat
     5641 cctctacgcc ggacgcatcg tggccggatc ttgcggccgc aaaaattaaa aatgaagttt
     5701 tgacggtatc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     5761 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     5821 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     5881 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     5941 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tccgcgcctt
     6001 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     6061 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     6121 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     6181 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     6241 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     6301 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tggagttcat tcagggcacc
     6361 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     6421 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     6481 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     6541 tgtctcttga tccactagat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6661 aagtgccacc tgcatcgatg gcccccgatg gtagtgtggg gtctccccat gcgagagtag
     6721 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     6781 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     6841 aa
//
