LOCUS       sequence-482751-        9100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLAM12 vector backbone containing Kva1 recombitron with MspRecT and 
            a donor in the ncRNA to target MSMEG_5894 gene of Mycobacterium 
            smegmatis mc2 155.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9100)
  AUTHORS   Gonzalez-Delgado A, Bonillo-Lopez L, Johnson MS, Knodlseder N, Ko 
            CC, Lekbach Y, Oh JH, Selvakumar H, Wold MC, Yu Z, Aragon V, 
            Gralnick JA, Guell M, Hatfull GF, Keitz BK, Koskella B, Mutalik VK, 
            van Pijkeren JP, Shipman SL
  TITLE     Genome editing of phylogenetically distinct bacteria using portable 
            retron-mediated recombineering
  JOURNAL   bioRxiv 2025.06.16.660010
REFERENCE   2  (bases 1 to 9100)
  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: "bioRxiv 
            2025.06.16.660010"
COMMENT     Sequence Label: pRECMYC-Kva1
FEATURES             Location/Qualifiers
     source          1..9100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(4578..5166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4658..4677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5497..6312)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6220..6239
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6304..6323
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(6473..6519)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggccggtttc ccagttgccc gacacgatca ggaaggccgg cacggcggcg gtgatcacgc
       61 cttcggccac cgcgacgaac ccgacggcgg gccccagtgc gtcgtgcccc aggccgagca
      121 gcaggaacag catgaaccac agcactgccc acagcagcca gatcaccccg aacgccgggt
      181 cggcctcggc ggtgaacgcg tgccacgagt agccgagtgc ggcgatcgcg acgaacagcg
      241 agaaccatcc gagaccttct ccgtcccagt cggtcacgtt gttgatggcc acccacaggt
      301 aggtgaagcc gaacaggtag agcccggagg ccgcgaagat cacggccgcg tctccgccgg
      361 actgcaggat cagcaccgta ggcgtcacca cctgcagtcc gccgacgaag aagttcagcg
      421 gtgttgcgcc tcgtgggctg atgcggccca gcagcatcag cccgtcgatg atgagcaccg
      481 cacccacgta gaagagcccg acgccaccca tgtcgctagt acctcggcag actcggtaga
      541 ccgggattgc ggcgcggggc ccgcgggcgg tccgcgcctg cgggaatcga cttcaccacc
      601 gtgggacttt cggcgctcgc ggacgccgcg tcgaatgcgc ggtgatgtcc cgcggtgaat
      661 gtcgtcagcc ctaccgaacc gaagacccgg cgcgccggcg tccggcactc cggacaggtc
      721 gccgccgcat cgcgctcgga gatcgcgcag gtgagatcga agggaccgca gtgggaacaa
      781 cggaatgtgt aggtcggcat ggtcacccct ttccattcac ccgaaatata tccagatgaa
      841 aataaaagtc aagggtgaac gcccgcgccg ggcgcgggtg gttacgaagt caaattcgag
      901 gtggtgtcga cgagttcgcg caactggtcg acgtccagcc cgcacgattc aaaagccgca
      961 cactcggcct cggctaccgc gtccaccagg cggttgcgca gcgtccgccc ccgcggggtc
     1021 aacgccacga gcacccgccg gcggtcatgc agatcggcag tccggtagat gatcccttgc
     1081 gacacaaggc gatcgacggt tctggtgaga gtcggacccg tgatcagcgt ggcctgcgcg
     1141 agatcgctca tgggcacggt gcgccgtccg gccaactcgt cgagcacgcg ccagtcctcg
     1201 atcttgagcg acgagtccgc gagcacgcgt tcgatcgccg actccgcgag gtgggtcaga
     1261 cgcaccaacg gcagcaggag gttggggcgg cgctgctgtc cggcggccat ggtgggtgtc
     1321 tcctcgtgtc gacggcgggg tctgttcaga atgaattatt tctgctagaa agctttgaag
     1381 cgcagactac acgggatcga cggtgatctg gagtgcagga cggcgaggtt gagttccgcg
     1441 tcgggctggt gattccgctt caagggccag cgggaatctt cgcgccttcg tgcgaggccg
     1501 tggccgagct cgcggccaag gaggtcaacg accgaggcgg cctgcagggc cgcaaggtca
     1561 ccatcgaggt gctcgacggc ggagcgcccg gcgacgacgt cgcccggacc gtcgccgacc
     1621 ggttgcgagg tcacggtctc gacgccgtga ccggctggca catctcggcc gtgcgcaacc
     1681 gcatctcccc ggtggtccgc gaccgcatcc cgtacgtcta cacctcgttg tacgagggcg
     1741 gtgaacgcac accgggcgtg ttctgcacag gcgagacacc gcagatccag atcgcgcccg
     1801 cgctcgcctg gctgcgcgac cacttcggca tccggtcctg gtgcctggtc ggcgatgact
     1861 acatctggcc gcgccgttcc gccgcggccg cccgcgcgta ctgccgagat ctggacctcg
     1921 agctcagacg ggagatctac gtcccgtacg gcaccgacga tttccgcgca cctgtccgca
     1981 aggccatcgc gtcgggggcg caggccgtgt tgatgctgct cgtcggccag gacgccgtgt
     2041 tgttcaaccg cgagttcgcg cgcgccggcg ggcacgaccg catggcgcgg ttcagcccgc
     2101 tgatggagga gaacatgctg ctggccagcg gcgccggctc caccgaaaac ctctatgtcg
     2161 ccgcggcgta cttcagttca ctggccaccg cgggtgccat ggacctgatg ggcagctacg
     2221 tcgcccgcta cggcgccgac gcgccaccgc tcaacgcgat ggccgaatcc tgttacgagg
     2281 gtctgcttgc gctcgaggcc atcttccagc gggcccactc ccccgagata ccggacctga
     2341 tggcatccgc gcacgatgtc gggttcgacg ggccccgggg cccgatgtgt atgcgcgaca
     2401 gtcaattcga ccagcaggtc tacatcgcca gcgccgacgg ctacgacttc gacatcctcg
     2461 acacgctggc gacgctcgac gcttgagacc gcgtcacttc tttatctaga tttaaagatc
     2521 tggtaccgcg gccgcgccac ggatgccacc acaggcacta caacggagtt cgccacgtac
     2581 atcaccacaa ccaccgattc tggcggtgag ctccccgata ttcagcggaa atggcttggt
     2641 atcgaccaag attcgtagaa ccccgtctcg tctggctggt attcaaaacg ggcgcaacga
     2701 aacacgcaac gagacaggca tggcccaaac cagaaaacta gcgtctacca ggacttttac
     2761 ctgtccgacc cgttgcaacg gaacccccca cggaaccccc gcgacacccg ctccccaatt
     2821 gcgttagaac agcggtggat tgtcggcttc gttgtgggcc ttttgagccg cttcctgttc
     2881 tgccgcacgc tctttcctcg cccgatagcc gagtcgctta acggtgtcca gatgcagccc
     2941 gaaatgtttg gccgtttgcg gccaagagtg gccctcgtcg tcgtgatagg cgcggatgcg
     3001 ttcgcggcgt gcagcctgct cggcgagcca ctcgctgcgt tcctgcgcca cgagccggac
     3061 gacgtggcgt tcggatagtc cggtgattcg agcgccttcg gcggcggtca cgcgccgctt
     3121 tttgcggaca gtcggctgcc ggttgtagcc gtcgctgtag ccgtcgctca tagcaatgcc
     3181 tccatggctg acgcggactt tgcgcgccgc gcaactgtgc tcgccgccgt gcgcgctgct
     3241 gcgcccttcc gcgagatggc cgactggcgc gcactgagtg tggcctcgta gaccacgatc
     3301 ccgtccgccc aaatgcgcga cttggttgtg atccaacgcc aaatgctgtt ggcgatggcg
     3361 cggacctcgc tgtccggtag cggtccggga cacacgtcgt tgcacggaaa ttcggcgttt
     3421 cgcgcgtggc actcggcata gatcgcgcgg ccgagtccgt ccacgttccg ggtcggcagg
     3481 tagatccgca tgagggcggg acgataggcc cacaacctga cggaatcgaa cagtgcgcaa
     3541 ttccgcccta gcggcgtcgg agccgctttg tacgtggtct gctgacgcca gcgcggcggt
     3601 ggcatgttcg cgccgagctc ggcctcgatg tggctgagtg tgtagagatc tgagtggagc
     3661 cattccgttt cccaggcgat gtggccgggg tttttggtca tgaggcctga gtaactgcgg
     3721 tcgccatcga cggcgcgccg aaggccttcg gcgcacgccg ccatgtatgc gagcggctta
     3781 cgccgcgcgt attcggtgcg tggaacaggg gcgttgagtg cccacactgc gtgtgcgtgg
     3841 ccgttggcgc gattgcccac gatcgcgttg ggcagcggat gggacccccg ggcgctgagc
     3901 gctcggagcg ctgcgtctgg atggtctacg tccacgacca gcaggtttgc cagcgctgtt
     3961 gggttcgcct cgatgtaccg gcggcctagg gccgacgcgc ggctttggcg gtagatcccc
     4021 tcgagcagat cgtcgcttgc cagcggccag tacggcagcc agagctgctc aaattcgtcg
     4081 gcgacgtggc tcacgcttgg tagtagacca cgattaatca ccggtgtatg gtccgacacg
     4141 agctccaagt cagatatttc gctgaggggc caccccacaa ctgcacactc ccccgctctc
     4201 ccgtcgagcc ctggtggtgg aacaccagcg acagccgagc acccccaacc acctgtacca
     4261 accaggagga acacatgcgt cgtttcgagg acgtttccgg gccgctgaga gccgctgtgg
     4321 cggccgtaca cgccgcctta gacccgttag accccctgcc gcctgaatgc gcgggtacga
     4381 gccacacagc gcccgaactt acggagctgg tgggctcacg cgttgcgctc ggtcgttcgg
     4441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4741 tttctcccct cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4981 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcatggaac tagtgaggtc
     5221 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     5281 agaaagtgag ggagccacgg ttggtgagag ctttgttgta ggtggaccag ttggtgattt
     5341 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     5401 tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat
     5461 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     5521 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5581 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5641 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     5701 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     5761 attatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     5821 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     5881 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     5941 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     6001 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     6061 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     6121 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat ggggcttccc
     6181 atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     6241 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     6301 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6361 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     6421 ttcccccccc cccgacgtca ggtggctagc tgatcaccgc ggccatgatg gcataaaacg
     6481 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag tcgatcgtac
     6541 gctagttgca gatcaaccac cgcgtcggaa ttcgccgggg cgctcatgcg ttgggcccgt
     6601 cgaacatgtc gatgacagcc ttcgcctcgt cgaacgtgag gtcggcggcg cgcgtggctt
     6661 tgacgccggc cgaatcggcg aggaacttga accaatcggc gtcgttgtac ttttcggcct
     6721 tctggatctc gccgaggcgc ttcagctgat cgggattggc catctgcacg tcgggcgccg
     6781 gcgcggtggc gcgctgcgcg cgttgtgcgc gcgacgactt cggtggcgta tcgacctcgt
     6841 gcggctcggc gatctcgccg tcgatgtagt cggccgtcgg cttgacggtc gacggcagcg
     6901 cgagcgcctg cgactgtgac aggtcggtgc ccgggcgatc gtcggcgcgg atcgccgcgt
     6961 cgagccgcgt cgtcttcggt gccagcttca gcacctgttt gagcgcggtc ttccgctcca
     7021 tccagcgctg cgggtccttg atgtcgcccg aggatccgac cttgccgcga cgcaattccc
     7081 tgatctcgtc ggcggtgaac acgtcccaca gcggctcagc gccggtcacc tcgacgatcg
     7141 cgtagaagta gaccggatta ccccgatcac ccttggcctt cacgtgtttc agcgtcggat
     7201 tgaggcccat ggtgtaatgg aattcgtcgt tcgcgccgac ccactgcgcg tcgatccggg
     7261 aggcgcgcgg gtgctgccag aacagtttga cgatgccctg gtagccgatg atcagctggc
     7321 actcgaccac gccgcgccgg gtgtcgcggt agggcacgag gtagcactcg ccgttgacac
     7381 cgggctcgag accgagcgcg gccgaggtca gcagcgcacc ggcaaatgac tcttgcgtgc
     7441 agtcgtcgag ggactgctta gcgatgccag cagctttcgc gttgcgcgac ttgcggatct
     7501 cggtttgcac gatccgcgcg atgcggtcgg ggtccatacc cttgggcact gcacgcgcca
     7561 gctgcggcac gagcacctgc agcacttgcg agatcgtctc gggtgccttg ggcgaatcct
     7621 gcttggtgac agcattttca gccatgattg gctttctcct ttctcccgcc gggatcatgc
     7681 aggtggcagc cgcttatgtt ataaaaaacc ccggtaaacc ggggttctgc tctttaccta
     7741 gaagggcacg gcctcctctg gcgccggggt gatggtgtcc agcggctcac acgggccccg
     7801 ccgggcgtga cgcacgacgg cgcaaggccg tggcataaat gaccacaatc aacaagttcc
     7861 ggtgatacac cagatgaagc agatacgcct ctatcgcgta aagagtatgt tatacactaa
     7921 cttaatgatt tgcatcaatc aagctgcttg tctcaagatt atggatcacc tgctctagcc
     7981 tcatatgaac ggtattagtc gttgtttttt tgcaggaccg caaacgagtc tttgcccagt
     8041 tgtttgagtt tgtcctcccc attgatctta atgagcgagt aatagtagtc cggttcaatg
     8101 ttcttgatat gggacagcat cccgaggaga tagggaatat ctttatgatt gattttatta
     8161 ttgctataca gatagaacag cgcacgaatc tttttgcgca ggaaccgacc cacgccgagg
     8221 tccccggtat tcgagatcac cacccccgta atccggcgcg agcgaccttt cccgataaat
     8281 ttggttttcg acgtattaat gatcagctcg ggtttttttt gcagcgagac caaattaatc
     8341 acgaaatttt tgacatcatt gagcttgtcg aacgaatcgg agctaaagga caggtcgtcc
     8401 gcataccgcg tgtagatcac atcattctct ttacagtagt taaacaattt ttcgtcgaaa
     8461 tcatacatga caatattgga gatcatcggg ctggagggcg ccccgataca caagcggagc
     8521 tttttctcgc cctttttgcg acgaaagaga tagtgggaaa tcacttcata ctcaaattgg
     8581 ttcaacttga tgtcattttt cgagataaag tagttcaggt cctgcggctt gatgctatag
     8641 aagaagttct cgaaatccat cttcaggata tactcatttt ttttgtgggg ttcggcatta
     8701 tccacgatcg acttttcctc gatgtaggcc gtggcacatt cgtgcactgg gagcagggac
     8761 aagaggctgt tcacaatggc ccgttggacc ttcttgacgt cgggcgacgg ctgggcaatt
     8821 tcgcgcagac ccttcccatt ttttttctta ataaagaaca ctttgtattt cttgggggct
     8881 ttaatataac cggagatgat ggtctttggg gtacaattca gcgtcttcga cagctcgtca
     8941 atatagctca tatgtggact ccctttctct tatcgggtgg tggccgcgag cgcggccggt
     9001 ggagggttgg tgactgacgg cgctgcgaga cggcgcccga tggggtatgc gagaacaact
     9061 gcggcaaaac cgatcacggc gatgaccgcg gcggggagcg
//
