LOCUS       sequence-367521-        6410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host range regulated expression of the DddA immunity 
            determinant (DddAI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6410)
  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 6410)
  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-dddAI
FEATURES             Location/Qualifiers
     source          1..6410
                     /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             473..505
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     terminator      539..585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(874..892)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1119..2201
                     /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(1138..1157)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2245..2250,2251..2267,2268..2273)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2462..2995
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            3122..3231
                     /note="incP origin of transfer"
     rep_origin      3623..4211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4112..4131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4365..4382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4397..4537
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4639..4830)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5348..6181)
                     /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 aaagatgtac gcagacgatt tcgacggcga
      121 aattgagatc gatgaggttg atagcttggt cgagttcctc agcagaagac ctgcatttga
      181 cgcaaataat tttgttttga cgtttgagga aagtggtttt cctcaattga atatttttgc
      241 aaaaaacgat atcgccgttg tttattatat ggacatcggc gagaatttcg tttcgaaagg
      301 caattctgcg tcaggtggta cggagaagtt ttacgagaat aagcttggcg gggaagtaga
      361 cctgtcgaaa gattgcgttg tgagtaaaga gcaaatgatt gaagcggcaa agcagttttt
      421 tgctacgaag caacggcccg agcaactgac atggagcgag ttgtaatcta gatatacaga
      481 tattgaaatg aatagattag gaaaatgagc tagagtcgac ctgcaggcat gcaagcttta
      541 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgatcgc ccttcccaac
      601 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
      661 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      721 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      781 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      841 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
      901 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
      961 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1021 aataaccctg ataaatgctt caataatatt aatggagcaa aacctttcgc ggtatggcat
     1081 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     1141 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     1201 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     1261 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     1321 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     1381 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     1441 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     1501 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     1561 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     1621 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     1681 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     1741 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     1801 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     1861 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     1921 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     1981 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     2041 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     2101 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2161 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     2221 agaatattac gcgtaattct cgaattgaca taagcctgtt cggttcgtaa actgtaatgc
     2281 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     2341 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg
     2401 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     2461 gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
     2521 tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat
     2581 gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca
     2641 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc
     2701 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga
     2761 gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca
     2821 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     2881 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     2941 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt
     3001 cgttcaagcc gagatcggct tcccggccga cgcgtcaatt ccgtgatagg tgggctgccc
     3061 ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta
     3121 gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca
     3181 gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc
     3241 cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg
     3301 aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg
     3361 gaaaacggaa ttggacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3421 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4321 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4381 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4441 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4501 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4561 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4621 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4681 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4741 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4801 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4861 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4921 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4981 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     5041 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     5101 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     5161 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     5221 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     5281 gatcatgcgc acccgtggaa attaattaat tgcagaaagg caggccgggc cgtggtggcc
     5341 acggcctcta ggccagatcc agcggcatct gggttagtcg agcgcgggcc gcttcccatg
     5401 tctcaccagg gcgagcctgt ttcgcgatct cagcatctga aatcttcccg gccttgcgct
     5461 tcgctggggc cttacccacc gccttggcgg gcttcttcgg tccaaaactg aacaacagat
     5521 gtgtgacctt gcgcccggtc tttcgctgcg cccactccac ctgtagcggg ctgtgctcgt
     5581 tgatctgcgt cacggctgga tcaagcactc gcaacttgaa gtccttgatc gagggatacc
     5641 ggccttccag ttgaaaccac tttcgcagct ggtcaatttc tatttcgcgc tggccgatgc
     5701 tgtcccattg catgagcagc tcgtaaagcc tgatcgcgtg ggtgctgtcc atcttggcca
     5761 cgtcagccaa ggcgtatttg gtgaactgtt tggtgagttc cgtcaggtac ggcagcatgt
     5821 ctttggtgaa cctgagttct acacggccct caccctcccg gtagatgatt gtttgcaccc
     5881 agccggtaat catcacactc ggtcttttcc ccttgccatt gggctcttgg gttaaccgga
     5941 cttcccgccg tttcaggcgc agggccgctt ctttgagctg gttgtaggaa gattcgatag
     6001 ggacacccgc catcgtcgct atgtcctccg ccgtcactga atacatcact tcatcggtga
     6061 caggctcgct cctcttcacc tggctaatac aggccagaac gatccgctgt tcctgaacac
     6121 tgaggcgata cgcggcctcg accagggcat tgcttttgta aaccattggg ggtgaggcca
     6181 cgttcgacat tccttgtgta taaggggaca ctgtatctgc gtcccacaat acaacaaatc
     6241 cgtcccttta caacaacaaa tccgtccctt cttaacaaca aatccgtccc ttaatggcaa
     6301 caaatccgtc cctttttaaa ctctacaggc cacggattac gtggcctgta gacgtcctaa
     6361 aaggtttaaa agggaaaagg aagaaaaggg tggaaacgca aaaaacgcac
//
