LOCUS       sequence-402539-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the gene codying for the Ssr recombinase from Pseudomonas 
            putida DOT-T1E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Pankratz D, Gomez NO, Nielsen A, Mustafayeva A, Gur M, 
            Arce-Rodriguez F, Nikel PI, Haussler S, Arce-Rodriguez A
  TITLE     An expanded CRISPR-Cas9-assisted recombineering toolkit for 
            engineering genetically intractable Pseudomonas aeruginosa isolates.
  JOURNAL   Nat Protoc. 2023 Oct 5. doi: 10.1038/s41596-023-00882-z.
  PUBMED    37798358
REFERENCE   2  (bases 1 to 6122)
  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"; journalName: "Nat Protoc.
            2023 Oct 5. doi: 10.1038/s41596-023-00882-z."
COMMENT     Sequence Label: pSH124-ssr
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..88
                     /gene="lacI (mutant)"
                     /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             89..1171
                     /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(108..127)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1403..1408,1409..1425,1426..1432)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1440..1456
                     /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)."
     terminator      2342..2436
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2509..2600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2601..2669,2670..3461)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3465..3504
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            3662..3770
                     /note="incP origin of transfer"
     CDS             complement(3831..4979)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      5372..5903
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6030..6116)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaatt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
       61 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      121 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      181 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
      241 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
      301 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
      361 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
      421 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
      481 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
      541 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
      601 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
      661 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
      721 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
      781 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
      841 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
      901 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
      961 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     1021 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     1081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg
     1201 aattgatctg gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     1261 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     1321 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     1381 gcaaatattc tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat
     1441 tgtgagcgga taacaatttc acaccctagg ccgcggccgc gcgaattcag aaggagaata
     1501 taccatgagc caagtagcca gggtcgaaac ccattcccag ccacaggctg tagccgccga
     1561 gtcggcgacc atcctgcaga tcatccagca agtcgcaatg tcccccaatg cagacatcga
     1621 caagatggag cgattgatgg cgatgcaccg ccagcaccaa gcacaacagg cgcagcaagc
     1681 gttcgatgcc gcgctagccg ccatgcagga agagcttccg gttatccgtg agcgcggcgc
     1741 aatcaaggac aagtacaaga acgtccaatc tacctacgcc ctatgggagg acatcaacga
     1801 agagttgaag cccatcctgg cgaagcacgg ctttgctctc accttccgaa ttccacgcac
     1861 cgacaaaggc atcgaggttg agggcgtgct tagtcatcga gacggccaca gggaaaccac
     1921 atcaatcctc ctgccagccg atgctaccgg cagcaagaat gccgtccagg ccgttgccag
     1981 ttctgtcagc tacgggaagc gctacacggc cggcgctctt ctgaacttca ccactactgg
     2041 cgaagacgat gacggccaag gagccgtacc gatgcaagtg cctgatgagc cggtcatcac
     2101 accgcgccaa gccgcccaac tcgatgggct gctgaaaaaa tgcagccaag tcctggtcga
     2161 caacttcacc gccaagtacg gctgcgctgc caacgtctac aaatccgagt tcgacgttgt
     2221 gctcgctcgc ctcaccaagt cggccagccg gccgcaggag taaggatcct ctagagtcga
     2281 cctgcaggca tgcaagcttg cggccgcgtc gtgactggga aaaccctggc gactagtctt
     2341 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     2401 ctcggttgcc gccgggcgtt ttttattggt gagaatccag gggtccccaa taattacgat
     2461 ttaaattagt agcccgccta atgagcgggc ttttttttaa ttcccctatt tgtttatttt
     2521 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2581 aatattgaaa aaggaagagt atgagcattc agcattttcg tgtggcgctg attccgtttt
     2641 ttgcggcgtt ttgcctgccg gtgtttgcgc atccggaaac cctggtgaaa gtgaaagatg
     2701 cggaagatca actgggtgcg cgcgtgggct atattgaact ggatctgaac agcggcaaaa
     2761 ttctggaatc ttttcgtccg gaagaacgtt ttccgatgat gagcaccttt aaagtgctgc
     2821 tgtgcggtgc ggttctgagc cgtgtggatg cgggccagga acaactgggc cgtcgtattc
     2881 attatagcca gaacgatctg gtggaatata gcccggtgac cgaaaaacat ctgaccgatg
     2941 gcatgaccgt gcgtgaactg tgcagcgcgg cgattaccat gagcgataac accgcggcga
     3001 acctgctgct gacgaccatt ggcggtccga aagaactgac cgcgtttctg cataacatgg
     3061 gcgatcatgt gacccgtctg gatcgttggg aaccggaact gaacgaagcg attccgaacg
     3121 atgaacgtga taccaccatg ccggcagcaa tggcgaccac cctgcgtaaa ctgctgacgg
     3181 gtgagctgct gaccctggca agccgccagc aactgattga ttggatggaa gcggataaag
     3241 tggcgggtcc gctgctgcgt agcgcgctgc cggctggctg gtttattgcg gataaaagcg
     3301 gtgcgggcga acgtggcagc cgtggcatta ttgcggcgct gggcccggat ggtaaaccga
     3361 gccgtattgt ggtgatttat accaccggca gccaggcgac gatggatgaa cgtaaccgtc
     3421 agattgcgga aattggcgcg agcctgatta aacattggta aaccgataca attaaaggct
     3481 ccttttggag cctttttttt tggacgaccc cggtccgcgc gttgtccttt tccgctgcat
     3541 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
     3601 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
     3661 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
     3721 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc
     3781 gcgatgcagg tggctgctga acccccagcc ggaactgacc ccacaaggcc ctagcgtttg
     3841 caatgcacca ggtcatcatt gacccaggcg tgttccacca ggccgctgcc tcgcaactct
     3901 tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac
     3961 atgaggcgga aggtttccag cttgagcggg tacggctccc ggtgcgagct gaaatagtcg
     4021 aacatccgtc gggccgtcgg cgacagcttg cggtacttct cccatatgaa tttcgtgtag
     4081 tggtcgccag caaacagcac gacgatttcc tcgtcgatca ggacctggca acgggacgtt
     4141 ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg acaccgattc caggtgccca
     4201 acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc
     4261 ttcgtgtaat accggccatt gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg
     4321 aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc
     4381 agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac gtccttgttg
     4441 acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg
     4501 aacagggcag agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca
     4561 atatcgaaca aggaaagctg catttccttg atctgctgct tcgtgtgttt cagcaacgcg
     4621 gcctgcttgg cttcgctgac ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg
     4681 gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt
     4741 tcgagacgac gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg gggagccagt
     4801 tgcacgctgt cgcgctcgat cttggccgta gcttgctgga ctatcgagcc gacggactgg
     4861 aaggtttcgc ggggcgcacg catgacggtg cggcttgcga tggtttcggc atcctcggcg
     4921 gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt
     4981 caccctcctt gcgggattgc cccggaatta attccccgga tcgatccgtc gatcttgatc
     5041 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     5101 caaccttacc agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg
     5161 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     5221 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     5281 gcggactggc tttctacgtg gctgccattt ttggggtgag gccgttcgcg gccgaggggc
     5341 gcagcccctg gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg
     5401 gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg
     5461 tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg
     5521 cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg
     5581 tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg
     5641 tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac
     5701 atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag
     5761 ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag
     5821 tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg
     5881 aggtatccac aacgccggcg gccctacatg gctctgctgt agtgagtggg ttgcgctccg
     5941 gcagcggtcc tgatcccccg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6001 acggcgcgcc cagctgtcta gggcggcgga tttgtcctac tcaggagagc gttcaccgac
     6061 aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     6121 ct
//
