LOCUS       sequence-482428-        9126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLAM12 vector backbone containing Ban1 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 9126)
  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 9126)
  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-Ban1
FEATURES             Location/Qualifiers
     source          1..9126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3541..3587
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3737..3756)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3748..4563
                     /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     complement(3821..3840)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4894..5482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5383..5402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcgaccag ttgcgcgaac tcgtcgacac cacctcgaat ttgacttcgt aaccacccgc
       61 gcccggcgcg ggcgttcacc cttgactttt attttcatct ggatatattt cgggtgaatg
      121 gaaaggggtg accatgccga cctacacatt ccgttgttcc cactgcggtc ccttcgatct
      181 cacctgcgcg atctccgagc gcgatgcggc ggcgacctgt ccggagtgcc ggacgccggc
      241 gcgccgggtc ttcggttcgg tagggctgac gacattcacc gcgggacatc accgcgcatt
      301 cgacgcggcg tccgcgagcg ccgaaagtcc cacggtggtg aagtcgattc ccgcaggcgc
      361 ggaccgcccg cgggccccgc gccgcaatcc cggtctaccg agtctgccga ggtactagcg
      421 acatgggtgg cgtcgggctc ttctacgtgg gtgcggtgct catcatcgac gggctgatgc
      481 tgctgggccg catcagccca cgaggcgcaa caccgctgaa cttcttcgtc ggcggactgc
      541 aggtggtgac gcctacggtg ctgatcctgc agtccggcgg agacgcggcc gtgatcttcg
      601 cggcctccgg gctctacctg ttcggcttca cctacctgtg ggtggccatc aacaacgtga
      661 ccgactggga cggagaaggt ctcggatggt tctcgctgtt cgtcgcgatc gccgcactcg
      721 gctactcgtg gcacgcgttc accgccgagg ccgacccggc gttcggggtg atctggctgc
      781 tgtgggcagt gctgtggttc atgctgttcc tgctgctcgg cctggggcac gacgcactgg
      841 ggcccgccgt cgggttcgtc gcggtggccg aaggcgtgat caccgccgcc gtgccggcct
      901 tcctgatcgt gtcgggcaac tgggaaaccg gcccgctccc cgccgcggtc atcgccgtga
      961 tcggttttgc cgcagttgtt ctcgcatacc ccatcgggcg ccgtctcgca gcgccgtcag
     1021 tcaccaaccc tccaccggcc gcgctcgcgg ccaccacccg ataagagaaa gggagtccac
     1081 atatgatcaa acaagtgctg cataacatca tctcgtccga tttgagcgaa gaagagaaaa
     1141 aatttttcat gcaatatgcc aaagatatcc actcgaacca ttgtccagtc atttttgata
     1201 ttaatcagat cctgcgcctc ttcgagttgc ccgacgacga cctggaaaaa ttcgtcattg
     1261 ataacaccaa ccactattat gtgtcgaaac ggaatgggga accccgtgaa gtgtggaaac
     1321 caaactataa gttgaaaacg attcaaaaat ggattctgaa aaatatcttg aataaaatcg
     1381 aggtctcggc gtgtgcgcat gggttcgtga agtccaagag cattttgacg aacgcggaag
     1441 tgcatcagca ccaagaacca ttctgggtgt tctccaccga catcaaagac ttcttcccct
     1501 cgattaaata taaagagatc aaaaaaatct ttattggtat tggctacagc gatgcggtgt
     1561 cggaggcctt ctccttgttg acgacggtga acaatgaact ggtccagggc tttcccacga
     1621 gcccgatgct gagcaatatc ttcttccgcg agatcgacga ggagtttcaa caactcgcca
     1681 gccggtataa tattcgttat tcgcgctatg ccgatgacat cacgttctcg ggcgtccaga
     1741 agaaggggta tctcaagttg gtcaaaaaga ttaaggagat tgtgtcgtgc gtgctgagca
     1801 aatacgattt cattatcaat aacaaaaaga cccggctgtt gaaggaccgt cataccaaaa
     1861 ttgtgaccgg cctggtgatt acgccgcagg gggtccgtat cccgcaaaag tacatccgcc
     1921 ggctgaataa agaaatctat tattgcaaaa aattcggcgt gaatgaacat ctcaagtatc
     1981 atggcctcat caccatcgcg aactacaagg ggtacctcat tggcctggcc cggttcatct
     2041 acatggtgga ccaaagcaaa ggtatcgagc tgatcaagaa aattcaggag ttggactggg
     2101 attaataccg ttcatatgag gctagagcag gtgatatgtt tagtggtgac tagactttta
     2161 gcaagtgcta atcgaagtaa gtatgtattt aattcttatt tcaaaaccat aatgatgatt
     2221 ctgcccaaaa ttgcagaacc atcctcacct ttaaattaag aatgcgccgt cgtgcgtcac
     2281 gcccggcggg gcccgtgtga gccgctggac accatcaccc cggcgccaga ggttaattta
     2341 aaggtgaagt agtcactaaa catgccacct gcatgatccg cggccgcacg gcgggagaaa
     2401 ggagaaagcc aatcatggct gaaaatgctg tcaccaagca ggattcgccc aaggcacccg
     2461 agacgatctc gcaagtgctg caggtgctcg tgccgcagct ggcgcgtgca gtgcccaagg
     2521 gtatggaccc cgaccgcatc gcgcggatcg tgcaaaccga gatccgcaag tcgcgcaacg
     2581 cgaaagctgc tggcatcgct aagcagtccc tcgacgactg cacgcaagag tcatttgccg
     2641 gtgcgctgct gacctcggcc gcgctcggtc tcgagcccgg tgtcaacggc gagtgctacc
     2701 tcgtgcccta ccgcgacacc cggcgcggcg tggtcgagtg ccagctgatc atcggctacc
     2761 agggcatcgt caaactgttc tggcagcacc cgcgcgcctc ccggatcgac gcgcagtggg
     2821 tcggcgcgaa cgacgaattc cattacacca tgggcctcaa tccgacgctg aaacacgtga
     2881 aggccaaggg tgatcggggt aatccggtct acttctacgc gatcgtcgag gtgaccggcg
     2941 ctgagccgct gtgggacgtg ttcaccgccg acgagatcag ggaattgcgt cgcggcaagg
     3001 tcggatcctc gggcgacatc aaggacccgc agcgctggat ggagcggaag accgcgctca
     3061 aacaggtgct gaagctggca ccgaagacga cgcggctcga cgcggcgatc cgcgccgacg
     3121 atcgcccggg caccgacctg tcacagtcgc aggcgctcgc gctgccgtcg accgtcaagc
     3181 cgacggccga ctacatcgac ggcgagatcg ccgagccgca cgaggtcgat acgccaccga
     3241 agtcgtcgcg cgcacaacgc gcgcagcgcg ccaccgcgcc ggcgcccgac gtgcagatgg
     3301 ccaatcccga tcagctgaag cgcctcggcg agatccagaa ggccgaaaag tacaacgacg
     3361 ccgattggtt caagttcctc gccgattcgg ccggcgtcaa agccacgcgc gccgccgacc
     3421 tcacgttcga cgaggcgaag gctgtcatcg acatgttcga cgggcccaac gcatgagcgc
     3481 cccggcgaat tccgacgcgg tggttgatct gcaactagcg tacgatcgac tgccaggcat
     3541 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatgcc atcatggccg
     3601 cggtgatcag ctagccacct gacgtcgggg gggggggaaa gccacgttgt gtctcaaaat
     3661 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     3721 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctcg
     3781 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     3841 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccca tgcgccagag
     3901 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     3961 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     4021 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     4081 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4141 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4201 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4261 ggctggcctg ttgaacaagt ctggaaagaa atgcataatc ttttgccatt ctcaccggat
     4321 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4381 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4441 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4501 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4561 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     4621 ggcggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     4681 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     4741 acaacaaagc tctcaccaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     4801 aggcctggta tgagtcagca acaccttctt cacgaggcag acctcactag ttccatgagc
     4861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5041 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5281 tgagctatga gaaagcgcca cgcttcccga ggggagaaag gcggacaggt atccggtaag
     5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5581 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcaacgc
     5641 gtgagcccac cagctccgta agttcgggcg ctgtgtggct cgtacccgcg cattcaggcg
     5701 gcagggggtc taacgggtct aaggcggcgt gtacggccgc cacagcggct ctcagcggcc
     5761 cggaaacgtc ctcgaaacga cgcatgtgtt cctcctggtt ggtacaggtg gttgggggtg
     5821 ctcggctgtc gctggtgttc caccaccagg gctcgacggg agagcggggg agtgtgcagt
     5881 tgtggggtgg cccctcagcg aaatatctga cttggagctc gtgtcggacc atacaccggt
     5941 gattaatcgt ggtctactac caagcgtgag ccacgtcgcc gacgaatttg agcagctctg
     6001 gctgccgtac tggccgctgg caagcgacga tctgctcgag gggatctacc gccaaagccg
     6061 cgcgtcggcc ctaggccgcc ggtacatcga ggcgaaccca acagcgctgg caaacctgct
     6121 ggtcgtggac gtagaccatc cagacgcagc gctccgagcg ctcagcgccc gggggtccca
     6181 tccgctgccc aacgcgatcg tgggcaatcg cgccaacggc cacgcacacg cagtgtgggc
     6241 actcaacgcc cctgttccac gcaccgaata cgcgcggcgt aagccgctcg catacatggc
     6301 ggcgtgcgcc gaaggccttc ggcgcgccgt cgatggcgac cgcagttact caggcctcat
     6361 gaccaaaaac cccggccaca tcgcctggga aacggaatgg ctccactcag atctctacac
     6421 actcagccac atcgaggccg agctcggcgc gaacatgcca ccgccgcgct ggcgtcagca
     6481 gaccacgtac aaagcggctc cgacgccgct agggcggaat tgcgcactgt tcgattccgt
     6541 caggttgtgg gcctatcgtc ccgccctcat gcggatctac ctgccgaccc ggaacgtgga
     6601 cggactcggc cgcgcgatct atgccgagtg ccacgcgcga aacgccgaat ttccgtgcaa
     6661 cgacgtgtgt cccggaccgc taccggacag cgaggtccgc gccatcgcca acagcatttg
     6721 gcgttggatc acaaccaagt cgcgcatttg ggcggacggg atcgtggtct acgaggccac
     6781 actcagtgcg cgccagtcgg ccatctcgcg gaagggcgca gcagcgcgca cggcggcgag
     6841 cacagttgcg cggcgcgcaa agtccgcgtc agccatggag gcattgctat gagcgacggc
     6901 tacagcgacg gctacaaccg gcagccgact gtccgcaaaa agcggcgcgt gaccgccgcc
     6961 gaaggcgctc gaatcaccgg actatccgaa cgccacgtcg tccggctcgt ggcgcaggaa
     7021 cgcagcgagt ggctcgccga gcaggctgca cgccgcgaac gcatccgcgc ctatcacgac
     7081 gacgagggcc actcttggcc gcaaacggcc aaacatttcg ggctgcatct ggacaccgtt
     7141 aagcgactcg gctatcgggc gaggaaagag cgtgcggcag aacaggaagc ggctcaaaag
     7201 gcccacaacg aagccgacaa tccaccgctg ttctaacgca attggggagc gggtgtcgcg
     7261 ggggttccgt ggggggttcc gttgcaacgg gtcggacagg taaaagtcct ggtagacgct
     7321 agttttctgg tttgggccat gcctgtctcg ttgcgtgttt cgttgcgccc gttttgaata
     7381 ccagccagac gagacggggt tctacgaatc ttggtcgata ccaagccatt tccgctgaat
     7441 atcggggagc tcaccgccag aatcggtggt tgtggtgatg tacgtggcga actccgttgt
     7501 agtgcctgtg gtggcatccg tggcgcggcc gcggtaccag atctttaaat ctagataaag
     7561 aagtgacgcg gtctcaagcg tcgagcgtcg ccagcgtgtc gaggatgtcg aagtcgtagc
     7621 cgtcggcgct ggcgatgtag acctgctggt cgaattgact gtcgcgcata cacatcgggc
     7681 cccggggccc gtcgaacccg acatcgtgcg cggatgccat caggtccggt atctcggggg
     7741 agtgggcccg ctggaagatg gcctcgagcg caagcagacc ctcgtaacag gattcggcca
     7801 tcgcgttgag cggtggcgcg tcggcgccgt agcgggcgac gtagctgccc atcaggtcca
     7861 tggcacccgc ggtggccagt gaactgaagt acgccgcggc gacatagagg ttttcggtgg
     7921 agccggcgcc gctggccagc agcatgttct cctccatcag cgggctgaac cgcgccatgc
     7981 ggtcgtgccc gccggcgcgc gcgaactcgc ggttgaacaa cacggcgtcc tggccgacga
     8041 gcagcatcaa cacggcctgc gcccccgacg cgatggcctt gcggacaggt gcgcggaaat
     8101 cgtcggtgcc gtacgggacg tagatctccc gtctgagctc gaggtccaga tctcggcagt
     8161 acgcgcgggc ggccgcggcg gaacggcgcg gccagatgta gtcatcgccg accaggcacc
     8221 aggaccggat gccgaagtgg tcgcgcagcc aggcgagcgc gggcgcgatc tggatctgcg
     8281 gtgtctcgcc tgtgcagaac acgcccggtg tgcgttcacc gccctcgtac aacgaggtgt
     8341 agacgtacgg gatgcggtcg cggaccaccg gggagatgcg gttgcgcacg gccgagatgt
     8401 gccagccggt cacggcgtcg agaccgtgac ctcgcaaccg gtcggcgacg gtccgggcga
     8461 cgtcgtcgcc gggcgctccg ccgtcgagca cctcgatggt gaccttgcgg ccctgcaggc
     8521 cgcctcggtc gttgacctcc ttggccgcga gctcggccac ggcctcgcac gaaggcgcga
     8581 agattcccgc tggcccttga agcggaatca ccagcccgac gcggaactca acctcgccgt
     8641 cctgcactcc agatcaccgt cgatcccgtg tagtctgcgc ttcaaagctt tctagcagaa
     8701 ataattcatt ctgaacagac cccgccgtcg acacgaggag acacccacca tggccgccgg
     8761 acagcagcgc cgccccaacc tcctgctgcc gttggtgcgt ctgacccacc tcgcggagtc
     8821 ggcgatcgaa cgcgtgctcg cggactcgtc gctcaagatc gaggactggc gcgtgctcga
     8881 cgagttggcc ggacggcgca ccgtgcccat gagcgatctc gcgcaggcca cgctgatcac
     8941 gggtccgact ctcaccagaa ccgtcgatcg ccttgtgtcg caagggatca tctaccggac
     9001 tgccgatctg catgaccgcc ggcgggtgct cgtggcgttg accccgcggg ggcggacgct
     9061 gcgcaaccgc ctggtggacg cggtagccga ggccgagtgt gcggcttttg aatcgtgcgg
     9121 gctgga
//
