LOCUS       sequence-250158-        3747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  UV5 promoter drives expression of Zif268-5alanine peptide fusion. 
            Pair with pGHUC w-ERBIN as a negative control (doesn't turn on 
            HIS3/GFP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3747)
  AUTHORS   Ichikawa DM, Corbi-Verge C, Shen MJ, Snider J, Wong V, Stagljar I, 
            Kim PM, Noyes MB
  TITLE     A Multireporter Bacterial 2-Hybrid Assay for the High-Throughput and
            Dynamic Assay of PDZ Domain-Peptide Interactions.
  JOURNAL   ACS Synth Biol. 2019 May 17;8(5):918-928. doi: 
            10.1021/acssynbio.8b00499. Epub 2019 Apr 18.
  PUBMED    30969105
REFERENCE   2  (bases 1 to 3747)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.8b00499"; journalName: "ACS Synth Biol"; date: 
            "2019-05-17- 17"; volume: "8"; issue: "5"; pages: "918-928"
COMMENT     Sequence Label: pB1H2 UV5 Zif268-5ala (2-hybrid bait plasmid)
FEATURES             Location/Qualifiers
     source          1..3747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(12..17,18..35,36..42)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    50..66
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     55..77
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             321..344
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      808..1263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(895..914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1105..1126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1289..1393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1394..1462,1463..2254)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1612..1631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2425..3013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2914..2933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3167..3184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3199..3339
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3265..3284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3425..3447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3441..3632)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaattccggg ctttacactt tatgcttccg gctcgtataa tgtgtcgact tgtgagcgga
       61 taacaatttc acacaggaaa cagctatgcc tcaacagcag caaatgcaac ctcccaattc
      121 aagtgcggac aacaaccctt tgcaacagca atcatcacaa aataccgtac caaacgtcct
      181 caaccaaatt aaccaaatct tttctccaga ggagcaacgc agcttattac aagaagccat
      241 cgaaacctgc aagaattttg aaaaaacaca attgtaagtc tcggctccaa gaaggagata
      301 tacccatggg tgcggccgcg gactacaagg atgacgacga caagttccgg acgggctcca
      361 agacaccgcc tcacggtacc gagcgcccat atgcttgccc tgtcgagtcc tgcgatcgcc
      421 gcttttctcg ctcggatgag cttacccgcc atatccgcat ccataccggt cagaagccct
      481 tccagtgtcg aatctgcatg cgtaacttca gtcgtagtga ccaccttacc acccacatcc
      541 gcacccacac aggcgagaag ccttttgcct gtgacatttg tgggaggaag tttgccagga
      601 gtgatgaacg caagaggcat accaaaatcc atctgcgcgg atccggcggt gggagcggag
      661 gcaccggcgc ggcggcggcg gcgtaatagg tctagagact agaaaaaggc cgacaagtcc
      721 cgctccgctg aagatcctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      781 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
      841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1261 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3181 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3241 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3301 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3361 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3421 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3481 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3541 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3601 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3661 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3721 gatgaacatg cccggttact ggaacgg
//
