LOCUS       sequence-471092-        2778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BsaI compatible gentamicin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2778)
  AUTHORS   VanDieren AJ, Barrick JE
  TITLE     UltraCAST: A Flexible All-In-One Suicide Vector for Modifying 
            Bacterial Genomes Using a CRISPR-Associated Transposon.
  JOURNAL   MicroPubl Biol. 2025 Aug 2;2025:10.17912/micropub.biology.001721. 
            doi: 10.17912/micropub.biology.001721. eCollection 2025.
  PUBMED    40827214
REFERENCE   2  (bases 1 to 2778)
  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: "MicroPubl 
            Biol."; date: "2025-08-2"; volume: 
            "2025:10.17912/micropub.biology.001721. doi"; pages: " 
            10.17912/micropub.biology.001721. eCollection 2025"
COMMENT     Sequence Label: pBTK1126
FEATURES             Location/Qualifiers
     source          1..2778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(814..819,820..836,837..842)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1031..1564
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     terminator      complement(1733..1760)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1833..1867)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1938..2597)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2512..2531
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtac gtttgagaag agaaaagaaa accgccgatc
      721 ctgtccaccg cattactgca aggtagtgga caagaccggc ggtcttaagt tttttggctg
      781 aaagtcggtc tcaaacgaca gcaatggatc gaattgacat aagcctgttc ggttcgtaaa
      841 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
      901 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct
      961 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     1021 agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     1081 aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt
     1141 caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg tagccaccta
     1201 ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat
     1261 cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc
     1321 caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggcgagca
     1381 ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct
     1441 tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta
     1501 tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac
     1561 ctaacaattc gttcaagccg agatcggctt cccggccgcg gagttgttcg gtaaattgtc
     1621 acaacgccgc ggccaattcg atctagaatt ccattgagta agtttttaag cacatcagct
     1681 tcaaaagcgc tctgaagttc ctagcggccg ctcggtgaaa ccgacttata aacgcagaaa
     1741 ggcccacccg aaggtgagcc gctgtgagac cggaaacaca gaaaaaagcc cacacctgac
     1801 agtgcgggct ttttttttcg accaaaggac caataaaaaa cgcccggcgg caaccgagcg
     1861 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     1921 gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     1981 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     2041 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     2101 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     2161 gaaacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     2221 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     2281 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     2341 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     2401 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     2461 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     2521 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     2581 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     2641 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc
     2701 aatcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2761 aaagatcaaa ggatcttc
//
