LOCUS       sequence-366192-        9008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ShCAST under control of Plac with pRO1600 origin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9008)
  AUTHORS   Trujillo Rodriguez L, Ellington AJ, Reisch CR, Chevrette MG
  TITLE     CRISPR-Associated Transposase for Targeted Mutagenesis in Diverse 
            Proteobacteria.
  JOURNAL   ACS Synth Biol. 2023 Jun 27. doi: 10.1021/acssynbio.3c00065.
  PUBMED    37368499
REFERENCE   2  (bases 1 to 9008)
  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: "ACS Synth 
            Biol. 2023 Jun 27. doi: 10.1021/acssynbio.3c00065."
COMMENT     Sequence Label: pRO16Sh5
FEATURES             Location/Qualifiers
     source          1..9008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      146..217
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(469..474,475..491,492..497)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             686..1219
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(1307..1323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1385..1736
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             1750..2583
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYCLSVQEQRIVLASISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMGSTHAIRLYELLTQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKARKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      2662..3250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3151..3170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            3320..3428
                     /note="incP origin of transfer"
     primer_bind     3616..3632
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(3694..3699,3700..3717,3718..3724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             3725..3736
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 aagtaaagca gctcaaacag gaattgtgat tgaggaggga aaacaaccta ttcgaggtag
       61 tccccacgac aaagcaaagg aattagcact ttctgcttac aatctccgcc taactaggcg
      121 aagttaagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
      181 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac gctttccagt cgggaaacct
      241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      301 gcgcatgcat aaaaactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
      361 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
      421 tttgcccatg gacgcacacc gtggaaacgg atgaaggcac gaacccagtt gacataagcc
      481 tgttcggttc gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
      541 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
      601 ttttttgtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
      661 tgtttgatgt tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg
      721 cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct
      781 cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg
      841 agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
      901 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
      961 ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
     1021 agtctccgga gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
     1081 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
     1141 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
     1201 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
     1261 gttcggtaaa ttgtcacaac gccgcaggtg gcacttttcg gggatcactg gccgtcgttt
     1321 tacaacagct ctgcagattt cgtgaattcg ccgctggtgc cgctggttgg acgccaaggg
     1381 tgaatccgcc tcgataccct gattactcgc ttcctgcgcc ctctcaggcg gcgatagggg
     1441 actggtaaaa cggggattgc ccagacgcct cccccgcccc ttcaggggca caaatgcggc
     1501 cccaacgggg ccacgtagtg gtgcgttttt tgcgtttcca cccttttctt ccttttccct
     1561 tttaaacctt ttaggacgtc tacaggccac gtaatccgtg gcctgtagag tttaaaaagg
     1621 gacggatttg ttgccattaa gggacggatt tgttgttaag aagggacgga tttgttgttg
     1681 taaagggacg gatttgttgt attgtgggac gctgatacag tgtcccctta tacacaagga
     1741 atgtcgaacg tggcctcacc cccaatggtt tacaaaagca atgccctggt cgaggccgcg
     1801 tattgcctca gtgttcagga acagcggatc gttctggcca gtattagcca ggtgaagagg
     1861 agcgagcctg tcaccgatga agtgatgtat tcagtgacgg cggaggacat agcgacgatg
     1921 gcgggtgtcc ctatcgaatc ttcctacaac cagctcaaag aagcggccct gcgcctgaaa
     1981 cggcgggaag tccggttaac ccaagagccc aatggcaagg ggaaaagacc gagtgtgatg
     2041 attaccggct gggtgcaaac aatcatctac cgggagggtg agggccgtgt agaactcagg
     2101 ttcaccaaag acatgctgcc gtatctgacg gaactcacca aacagttcac caaatacgcc
     2161 ttggctgacg tggccaagat gggcagcacc cacgcgatca ggctttacga gctgctcacg
     2221 caatgggaca gcatcggcca gcgtgaaata gaaattgacc agctgcgaaa gtggtttcaa
     2281 ctggaaggcc ggtatccctc gatcaaggac ttcaagttgc gagtgcttga tccagccgtg
     2341 acgcagatca acgagcacag cccgctacag gtggagtggg cgcagcgaaa gaccgggcgc
     2401 aaggtcacac atctgttgtt cagttttgga ccgaagaagc ccgccaaggc ggtgggtaag
     2461 gcccctgcga agcgcaaggc caggaagatt tcagatgctg agatcgcgaa acaggctcgc
     2521 cctggtgaga catgggaagc ggcccgcgct cgactaaccc agatgccgct ggatctggcc
     2581 tagaggccat ccatcacact ggcggccgct cgagtcgttc cactgagcgt cagaccccgt
     2641 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2701 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2761 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2821 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2881 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2941 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3001 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3061 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3301 tgctcacaag ctagcttatc ggccagcctc gcagagcagg attcccgttg agcaccgcca
     3361 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta
     3421 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa
     3481 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga
     3541 agcagggtta tgcagcggaa agtatacctt aaggaatccc catgttcttt cctgcgttcc
     3601 ccgaaaagtg ccaccgtaaa acgacggcca gtgagctggc cttttgctca acgccaagct
     3661 ccaaaacgat ctcaagaaga tcatcttatt aattttacac tttatgcttc cggctcgtat
     3721 gttgaaagag gagaaaggat ctatgaacag tcagcaaaat cctgatttag ctgttcatcc
     3781 cttggcaatt cctatggaag gcttactagg agaaagtgct acaactcttg agaagaatgt
     3841 aattgccaca caactctcag aggaagccca agtaaagcta gaggtaatcc aaagtttact
     3901 ggaaccctgc gatcgcacaa cttatgggca aaagttgcgg gaagcagcag agaaactaaa
     3961 tgtatcgttg cgaacggtac aaaggttggt gaaaaactgg gaacaagatg gcttagtcgg
     4021 actcactcaa acaagtaggg ctgataaagg aaaacaccgc attggtgagt tttgggaaaa
     4081 cttcattacc aaaacctaca aggagggtaa caagggaagt aaacgtatga cccctaaaca
     4141 agttgctctc agagtcgagg ctaaagcccg tgaattaaaa gactctaagc cgcccaatta
     4201 caaaaccgtg ttacgggtat tagcacccat tttggaaaag caacaaaaag ccaagagtat
     4261 ccgcagtcct ggttggagag gaactacgct ttcggttaaa acccgtgaag gaaaagattt
     4321 atcggttgat tacagtaacc atgtttggca atgtgaccat acccgcgtgg atgtgttgct
     4381 ggtagatcaa catggtgaaa ttttaagtcg tccctggcta acaacagtaa ttgatactta
     4441 ctctcgttgc attatgggta tcaacttggg ctttgatgca cccagttctg gggtagtagc
     4501 attagcgtta cgccatgcaa ttctaccaaa gcgttacggt tccgagtaca aactgcattg
     4561 tgagtgggga acctatggaa aaccagaaca tttttatact gatggcggta aagactttcg
     4621 ctctaaccac ttgagtcaga ttggggcgca attgggattt gtctgtcatt tacgcgatcg
     4681 cccttctgaa ggtggagtag tagaacgtcc cttcaaaaca ttaaatgacc aactattttc
     4741 aacgcttcct gggtacaccg gatctaatgt gcaggaacgc ccagaagatg cagagaagga
     4801 cgcaagactt actttgcgag aactagaaca gttacttgtg cgttacatcg tagatcgtta
     4861 caaccaaagt attgatgcgc ggatgggcga ccaaacgcgc tttgagcgtt gggaagcagg
     4921 attgcctaca gtgccagtac caataccaga acgagatttg gatatttgtt taatgaagca
     4981 gtcacggcgc actgtgcaaa gaggtggttg tttgcagttt cagaatttaa tgtatcgggg
     5041 ggaatatttg gcaggttatg ccggagaaac tgtcaactta aggtttgacc ccagagacat
     5101 tacaacaatt ttggtttatc gccaggaaaa caatcaggaa gtatttctga ctcgcgctca
     5161 cgctcaaggt ttggagacag agcaactggc attagatgag gctgaggcag caagtcgcag
     5221 actccgtacc gcagggaaaa ctatcagtaa ccaatcatta ttgcaagaag ttgttgaccg
     5281 cgatgctctt gtcgctacca agaaaagccg taaggagcgt caaaaattgg aacagactgt
     5341 tttgcgatct gctgctgttg atgaaagtaa tagagaatcc ttgccttctc aaatagttga
     5401 accagatgaa gtggaatcta cagaaacggt tcactctcaa tacgaagaca ttgaggtgtg
     5461 ggactatgaa caacttcgtg aagaatatgg gttttaaaca atgacagaag ctcaggcgat
     5521 cgccaagcag ttgggtgggg taaaaccgga tgatgagtgg ttacaagctg aaattgctcg
     5581 tctcaagggt aagagcattg tgcctttaca gcaggtaaaa actctccatg attggttaga
     5641 tggcaagcgc aaggcaagaa aatcttgccg agtagttggg gaatcgagaa ctggcaagac
     5701 agttgcttgt gatgcctaca gatacaggca caaacctcag caggaagctg gacgacctcc
     5761 aactgtgcct gtcgtttata ttcgacctca ccaaaaatgt ggccccaagg atttgtttaa
     5821 aaagattact gagtacctca agtatcgggt aacaaaaggg actgtatctg attttcgaga
     5881 taggacgata gaagtactca agggttgtgg cgtagagatg ctaattattg atgaagctga
     5941 ccgtctcaag cctgaaactt ttgctgatgt gcgagatatt gccgaagatt taggaattgc
     6001 tgtggtactg gtaggaacag accgtttgga tgcggtaatt aagcgggatg agcaggttct
     6061 cgaacgcttt cgggcgcatc ttcgctttgg taaattgtcg ggagaggatt ttaagaacac
     6121 cgtagaaatg tgggaacaaa tggttttgaa actgccagta tcttctaatc taaagagcaa
     6181 ggagatgcta cggattctca cgtcagcaac tgaaggctac attggtcgcc ttgatgagat
     6241 tcttagggaa gctgcaattc gttccttatc aagaggattg aagaagattg acaaggctgt
     6301 tttacaggaa gtagctaagg agtacaaatg atagaagcac cagatgttaa accttggcta
     6361 ttcttgatta aaccctatga aggggaaagc ctgagccact ttcttggcag gttcagacgt
     6421 gccaaccatt tatccgcaag tggattgggt actttggcag gaattggtgc tatagtggca
     6481 cgttgggaaa gatttcattt taatcctcgc cctagtcagc aagaattgga agcgatcgca
     6541 tctgtagtag aagtggatgc tcaaaggtta gcccagatgt taccgcctgc tggagtggga
     6601 atgcagcatg agccaattcg cttgtgtggg gcttgttatg ccgagtcgcc ttgtcaccga
     6661 attgaatggc agtacaagtc ggtgtggaag tgcgatcgcc atcaactcaa gattttagca
     6721 aagtgtccaa actgtcaagc accttttaaa atgcctgcgc tgtgggagga tgggtgctgt
     6781 cacagatgta ggatgccgtt tgcagaaatg gcaaagctac agaaggtttg atgataaaac
     6841 cagaaaaagg tgtgaaatta actaagtccc tgaattgatc tggttgtcca aaaaatttgt
     6901 gcgatcgcat ggcaagatta ttcctactat tgatgtggcg tgagtcggat aacttgctct
     6961 aatgggatcc cggttaatac caatagatca gatatgagta ataaacatta cgacctttac
     7021 ctgtggtatt cactgttggc acgacaacac ctttatcgcg ccaatactgt agctgacgac
     7081 gcgaacaatt agtgatttga gccgcttctg tgcttgtgta gaattttcct tccatagaaa
     7141 tcatcctaca gaagtagttt tttaaattta cattttttaa aagcacatta tgtattcatg
     7201 tgatattatt tttttatgag tcaaataact attcaagctc gacttatttc ctttgaatca
     7261 aaccgccaac aactctggaa gttgatggca gatttaaaca cgccgttaat taacgaactg
     7321 ctttgccagt taggtcaaca ccccgacttc gagaagtggc aacaaaaggg taaactcccg
     7381 tctaccgttg tgagccagtt atgtcaacct ctcaaaactg accctcgctt tgcaggtcag
     7441 cccagccgtt tatatatgtc ggcaattcat attgtggact acatctacaa gtcctggctg
     7501 gctatacaga aacggcttca acagcagcta gatggaaaga cgcgctggct agaaatgctc
     7561 aatagcgatg ctgaattagt agaacttagt ggtgacactt tagaggctat tcgtgtcaaa
     7621 gctgctgaaa ttttggcaat agctatgcca gcatctgagt cagatagcgc ttcacctaaa
     7681 gggaaaaaag gtaaaaagga gaaaaaaccc tcatcttcta gccctaagcg tagtttatcc
     7741 aagacattat ttgacgctta ccaagaaacg gaagatatca agagccgtag cgccatcagc
     7801 tacctgttaa aaaatggctg caaacttact gacaaagaag aagattcaga aaaatttgct
     7861 aaacgtcgtc gtcaagttga aatccaaatt caaaggctta ccgaaaagtt aataagtcgg
     7921 atgcctaaag gtcgagattt gaccaatgct aaatggttgg agacactctt gactgctaca
     7981 accactgttg ctgaagacaa cgcccaagcc aaacgctggc aggatattct gttaactcga
     8041 tcaagttctc tcccattccc ccttgttttt gaaaccaacg aggatatggt ttggtcaaag
     8101 aatcaaaagg gtaggctgtg tgttcacttc aatggcttaa gcgatttaat ttttgaggtg
     8161 tactgcggca atcgtcaact tcactggttt caacgcttcc tagaagacca acagactaaa
     8221 cgcaaaagca aaaatcagca ttctagcggc ttgttcacac tcagaaatgg tcatctagtt
     8281 tggcttgaag gtgagggtaa aggggaacct tggaatcttc accacttgac cctttactgc
     8341 tgtgttgaca atcgcttgtg gacagaggag ggaacagaaa tcgttcgcca agagaaagca
     8401 gatgaaatta ctaaattcat cacaaacatg aagaagaaaa gcgatctaag cgatacacag
     8461 caagctttga ttcaacgtaa acaatcaaca cttactcgaa taaacaattc ctttgagcgt
     8521 cctagccaac ccctttatca aggtcaatca cacattttgg ttggagtaag cctgggacta
     8581 gaaaaacctg ccacagtagc agtagtagat gcgatcgcca acaaagtctt ggcttaccgg
     8641 agtattaaac aattacttgg cgacaattac gaactgctaa atcgccagag acgacaacag
     8701 cagtacctat ctcacgaacg ccacaaagca caaaaaaact tctctcccaa tcaatttgga
     8761 gcatctgagt tagggcaaca tatagacaga ttattagcta aagcaattgt agcgttagcg
     8821 agaacctaca aagctggcag tattgtcttg cccaagttag gggatatgcg ggaggttgtc
     8881 caaagtgaaa ttcaagctat agcagaacaa aaatttcccg gttatattga aggtcagcaa
     8941 aaatatgcca aacagtaccg ggttaatgtt catcggtgga gctacggcag attaattcaa
     9001 agcattca
//
