LOCUS       sequence-367522-        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host range inducible expression of uracil-DNA glycosylase 
            inhibitor (UGI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  AUTHORS   Gallagher LA, Velazquez E, Peterson SB, Charity JC, Radey MC, 
            Gebhardt MJ, Hsu F, Shull LM, Cutler KJ, Macareno K, de Moraes MH, 
            Penewit KM, Kim J, Andrade PA, LaFramboise T, Salipante SJ, Reniere 
            ML, de Lorenzo V, Wiggins PA, Dove SL, Mougous JD
  TITLE     Genome-wide protein-DNA interaction site mapping in bacteria using a
            double-stranded DNA-specific cytosine deaminase.
  JOURNAL   Nat Microbiol. 2022 Jun;7(6):844-855. doi: 
            10.1038/s41564-022-01133-9. Epub 2022 Jun 1.
  PUBMED    35650286
REFERENCE   2  (bases 1 to 6293)
  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: 
            "10.1038/s41564-022-01133-9"; journalName: "Nat Microbiol"; date: 
            "2022-06"; volume: "7"; issue: "6"; pages: "844-855"
COMMENT     Sequence Label: pPSV39-ugi
FEATURES             Location/Qualifiers
     source          1..6293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(11..16,17..34,35..41)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    49..65
                     /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)."
     CDS             95..349
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             356..388
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     terminator      422..468
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(757..775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1002..2084
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1021..1040)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2128..2133,2134..2150,2151..2156)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2345..2878
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            3005..3114
                     /note="incP origin of transfer"
     rep_origin      3506..4094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3995..4014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4248..4265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4280..4420
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4522..4713)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5231..6064)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
ORIGIN
        1 cactacgtgc tttacacttt atgcttccgg ctcgtataat gtgtggaatt gtgagcggat
       61 aacaatttca gaattcgagc tcacgggagg aaagatgacg aatctcagcg acatcatcga
      121 gaaagaaacg gggaaacaac tggtgatcca ggaaagcatc ctgatgctcc ccgaggaggt
      181 cgaggaagtc atcggcaata aaccggagtc ggatatcctg gtccataccg cgtatgacga
      241 aagcacggac gagaacgtca tgctcctcac gtcggatgcg ccggaatata aaccgtgggc
      301 cctcgtgatc caagatagca atggcgagaa caaaatcaag atgctctaat ctagatatac
      361 agatattgaa atgaatagat taggaaaatg agctagagtc gacctgcagg catgcaagct
      421 ttagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgat cgcccttccc
      481 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
      541 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
      601 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      721 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      901 gacaataacc ctgataaatg cttcaataat attaatggag caaaaccttt cgcggtatgg
      961 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     1021 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     1081 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     1141 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     1201 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     1261 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     1321 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     1381 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     1441 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     1501 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     1561 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     1621 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     1681 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     1741 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     1801 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     1861 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     1921 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     1981 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2041 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2101 gtgagaatat tacgcgtaat tctcgaattg acataagcct gttcggttcg taaactgtaa
     2161 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     2221 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca gtctatgcct
     2281 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     2341 aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     2401 aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc aagtcaaatc
     2461 catgcgggct gctcttgatc ttttcggtcg tgagttcgga gacgtagcca cctactccca
     2521 acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat tcatcgcgct
     2581 tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt
     2641 tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggcg agcaccggag
     2701 gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg cgcttggtgc
     2761 ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc tctatacaaa
     2821 gttgggcata cgggaagaag tgatgcactt tgatatcgac ccaagtaccg ccacctaaca
     2881 attcgttcaa gccgagatcg gcttcccggc cgacgcgtca attccgtgat aggtgggctg
     2941 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     3001 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     3061 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     3121 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     3181 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     3241 gcggaaaacg gaattggacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3301 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4141 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4201 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4261 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4321 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     4381 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     4441 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4501 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     4561 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     4621 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     4681 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     4741 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     4801 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     4861 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     4921 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     4981 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     5041 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     5101 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     5161 cacgatcatg cgcacccgtg gaaattaatt aattgcagaa aggcaggccg ggccgtggtg
     5221 gccacggcct ctaggccaga tccagcggca tctgggttag tcgagcgcgg gccgcttccc
     5281 atgtctcacc agggcgagcc tgtttcgcga tctcagcatc tgaaatcttc ccggccttgc
     5341 gcttcgctgg ggccttaccc accgccttgg cgggcttctt cggtccaaaa ctgaacaaca
     5401 gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc cacctgtagc gggctgtgct
     5461 cgttgatctg cgtcacggct ggatcaagca ctcgcaactt gaagtccttg atcgagggat
     5521 accggccttc cagttgaaac cactttcgca gctggtcaat ttctatttcg cgctggccga
     5581 tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc gtgggtgctg tccatcttgg
     5641 ccacgtcagc caaggcgtat ttggtgaact gtttggtgag ttccgtcagg tacggcagca
     5701 tgtctttggt gaacctgagt tctacacggc cctcaccctc ccggtagatg attgtttgca
     5761 cccagccggt aatcatcaca ctcggtcttt tccccttgcc attgggctct tgggttaacc
     5821 ggacttcccg ccgtttcagg cgcagggccg cttctttgag ctggttgtag gaagattcga
     5881 tagggacacc cgccatcgtc gctatgtcct ccgccgtcac tgaatacatc acttcatcgg
     5941 tgacaggctc gctcctcttc acctggctaa tacaggccag aacgatccgc tgttcctgaa
     6001 cactgaggcg atacgcggcc tcgaccaggg cattgctttt gtaaaccatt gggggtgagg
     6061 ccacgttcga cattccttgt gtataagggg acactgtatc tgcgtcccac aatacaacaa
     6121 atccgtccct ttacaacaac aaatccgtcc cttcttaaca acaaatccgt cccttaatgg
     6181 caacaaatcc gtcccttttt aaactctaca ggccacggat tacgtggcct gtagacgtcc
     6241 taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac gcaaaaaacg cac
//
