LOCUS       sequence-318586-        3453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mini-transposon donor plasmid for V. cholerae CAST system. pUC19 
            vector backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3453)
  AUTHORS   Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH
  TITLE     CRISPR RNA-guided integrases for high-efficiency, multiplexed 
            bacterial genome engineering.
  JOURNAL   Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 
            10.1038/s41587-020-00745-y.
  PUBMED    33230293
REFERENCE   2  (bases 1 to 3453)
  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 
            Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 
            10.1038/s41587-020-00745-y."
COMMENT     Sequence Label: pSL1119 (pDonor)
FEATURES             Location/Qualifiers
     source          1..3453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             372..1031
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(438..457)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(1231..1248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1231..1247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1240..1262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1456..1475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1575..1597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1635..1653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1721..1825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1826..1894,1895..2686)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2044..2063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2857..3445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3346..3365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
       61 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      121 tcgccgcagc cgaactaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      181 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcc caagctttgt
      241 tgatacaacc ataaaatgat aattacaccc ataaattgat aattatcaca cccataaatt
      301 gatattgcct cttcatggtc taaacttcag taagtttacg acattttcct cgaggtcatt
      361 tccggggatc catggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
      421 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
      481 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
      541 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggagttc cgtatggcaa
      601 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
      661 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
      721 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
      781 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
      841 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcactatg ggcaaatatt
      901 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
      961 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1021 gcggggcgta atttttttaa ggcagttatt ggtgcccttc tagagtcctt actgcagtag
     1081 ttttgctgaa atactcgatt cacaaaaata tcaacttatg gttgttttgt gagatatcaa
     1141 tatatggttg ttttgtggtt aagttgctga ttataaataa ttattaaata tcactttatg
     1201 gttgcatcaa caggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     1261 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     1321 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1381 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     1441 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     1501 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     1561 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     1621 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1681 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1741 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1801 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1861 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1921 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1981 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2041 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2101 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2161 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2221 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2281 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2341 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2401 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2461 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2521 atctggagcc ggtgagcgtg gctctcgcgg tatcattgca gcactggggc cagatggtaa
     2581 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2641 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2701 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2761 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2821 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2881 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2941 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3001 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3061 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3121 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3181 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3241 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3301 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3361 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3421 gtcagggggg cggagcctat ggaaaaacgc cag
//
