LOCUS       sequence-482753-        9505 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  pLAM12 vector backbone containing Tsp1 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 9505)
  TITLE     Shipman lab plasmids 2025
REFERENCE   2  (bases 1 to 9505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRECMYC-Tsp1
FEATURES             Location/Qualifiers
     source          1..9505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     212..231
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(381..427)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(7991..8579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8071..8090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8910..9505,1..220))
                     /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"
ORIGIN
        1 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
       61 gaaacaactc tggcgcatgg ggcttcccat acaagcgata gattgtcgca cctgattgcc
      121 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
      181 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
      241 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
      301 atcagagatt ttgagacaca acgtggcttt cccccccccc cgacgtcagg tggctagctg
      361 atcaccgcgg ccatgatggc ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
      421 ttatttgatg cctggcagtc gatcgtacgc tagttgcaga tcaaccaccg cgtcggaatt
      481 cgccggggcg ctcatgcgtt gggcccgtcg aacatgtcga tgacagcctt cgcctcgtcg
      541 aacgtgaggt cggcggcgcg cgtggctttg acgccggccg aatcggcgag gaacttgaac
      601 caatcggcgt cgttgtactt ttcggccttc tggatctcgc cgaggcgctt cagctgatcg
      661 ggattggcca tctgcacgtc gggcgccggc gcggtggcgc gctgcgcgcg ttgtgcgcgc
      721 gacgacttcg gtggcgtatc gacctcgtgc ggctcggcga tctcgccgtc gatgtagtcg
      781 gccgtcggct tgacggtcga cggcagcgcg agcgcctgcg actgtgacag gtcggtgccc
      841 gggcgatcgt cggcgcggat cgccgcgtcg agccgcgtcg tcttcggtgc cagcttcagc
      901 acctgtttga gcgcggtctt ccgctccatc cagcgctgcg ggtccttgat gtcgcccgag
      961 gatccgacct tgccgcgacg caattccctg atctcgtcgg cggtgaacac gtcccacagc
     1021 ggctcagcgc cggtcacctc gacgatcgcg tagaagtaga ccggattacc ccgatcaccc
     1081 ttggccttca cgtgtttcag cgtcggattg aggcccatgg tgtaatggaa ttcgtcgttc
     1141 gcgccgaccc actgcgcgtc gatccgggag gcgcgcgggt gctgccagaa cagtttgacg
     1201 atgccctggt agccgatgat cagctggcac tcgaccacgc cgcgccgggt gtcgcggtag
     1261 ggcacgaggt agcactcgcc gttgacaccg ggctcgagac cgagcgcggc cgaggtcagc
     1321 agcgcaccgg caaatgactc ttgcgtgcag tcgtcgaggg actgcttagc gatgccagca
     1381 gctttcgcgt tgcgcgactt gcggatctcg gtttgcacga tccgcgcgat gcggtcgggg
     1441 tccataccct tgggcactgc acgcgccagc tgcggcacga gcacctgcag cacttgcgag
     1501 atcgtctcgg gtgccttggg cgaatcctgc ttggtgacag cattttcagc catgattggc
     1561 tttctccttt ctcccgccgg gatcatgcag gtggctcacg cgcatcgaag tgcaaattac
     1621 cccttcgtaa aggtctcagg actagactgc tccctctggc gccggggtga tggtgtccag
     1681 cggctcacac gggccccgcc gggcgtgacg cacgacggcg cagagcaggc tagtgatgca
     1741 tggttagact aagcacaagg acgacgaacc atcgtccggt ttgcaaggtt gtttcgcgta
     1801 gcttacagat gcgcgtgaat cacctgctct agcctcatat gaacggtact agctgcggcc
     1861 gttccgcacg tacttatttt taatgcgttg cacttgttgt tggaattcgg cgcctttctc
     1921 tggattcacc atcgcgacaa aattcgcgaa cccaccaatg gcggcgatga gattatcgga
     1981 gttgccccag tgcttaccct cgagcccgtc tttttcgatc tggaacaacg tggcacggaa
     2041 ccgtttcagg gtcttattgt cgacattcgg cttatcattg acgaccacac ccgtcacctc
     2101 ttgctgccgg gatttgcgca ggatacgggt cttctcttga ttaatcgtga aaccctcgtg
     2161 ggtgacgatg ctctcggtcc ccttcaagat attacagata tggcgcaagg agtcccccga
     2221 ggccgaaaac gtcaagtcgt ccgcgtaacg ggtgtaggta aagccgagtt tctcggccat
     2281 ggccgtcaac cgacgatcca aacggcggca caagagattg gtaatggctg gggacgcggg
     2341 ggacccttgt ggcaaatgcc gttcatccac cgccacgtag tacgtcttac catcaatctc
     2401 gagcgtttcc acttcggccg ccgtgctgac caggccaaag atcgtcgccg ccgcttcgga
     2461 gtaaccaaag gagtggaaga ggcccttcac acgcttgtag ctaatgcttg gaaaaaaatt
     2521 cttgaggtca aagttaataa tcacatccgc ccccacatgc ggcgtcgcgt tggtgacaat
     2581 ggagcgatca cggcggaagc cgtgcgccgc atcatgcaat tcgagcttct ccaggatatt
     2641 cgccaagatc cagtgctgcg cctgtttcaa gttgggcatc ggggccgaaa tgatacgctc
     2701 accccccgtc ttttttggca ttttgaaccg gatataatgg gagacggtcg acgtcttgcg
     2761 attaaaggcc aaaaaccgca acttgccgat cgtgatcccc atcgccgccg cgatttgctc
     2821 ggcattatta cacagtggca gattgtagct gtgcaaacgt tcttcgttac cttgcgtgtt
     2881 gttgaggccg gcgctgacat cctcgcccag atagatgata tcttgttgtt gtttctgctt
     2941 ccacgcttcc gcccgcgagc gccgctcttg ctcgcgacgt tccttcgtct cctgccgttt
     3001 gcgtttgctc tcctccaacc gttgcttcag gagccgtttg cggagttctt tttcatcttg
     3061 cagtttacgg gactcttgac gcaattcatt gagttcgcga cggatttcac ccgcgcggcg
     3121 aatctcgtcg gcggggtctt gtggcatctc accctgggcg ctccaaaaac cgtagcggat
     3181 catctcctcc aggataaact cttccttgcc gacttgacga atgcgatcgt acaattcctg
     3241 gcgcgtacgt ggttggtccg acatatgtgg actccctttc tcttatcggg tggtggccgc
     3301 gagcgcggcc ggtggagggt tggtgactga cggcgctgcg agacggcgcc cgatggggta
     3361 tgcgagaaca actgcggcaa aaccgatcac ggcgatgacc gcggcgggga gcgggccggt
     3421 ttcccagttg cccgacacga tcaggaaggc cggcacggcg gcggtgatca cgccttcggc
     3481 caccgcgacg aacccgacgg cgggccccag tgcgtcgtgc cccaggccga gcagcaggaa
     3541 cagcatgaac cacagcactg cccacagcag ccagatcacc ccgaacgccg ggtcggcctc
     3601 ggcggtgaac gcgtgccacg agtagccgag tgcggcgatc gcgacgaaca gcgagaacca
     3661 tccgagacct tctccgtccc agtcggtcac gttgttgatg gccacccaca ggtaggtgaa
     3721 gccgaacagg tagagcccgg aggccgcgaa gatcacggcc gcgtctccgc cggactgcag
     3781 gatcagcacc gtaggcgtca ccacctgcag tccgccgacg aagaagttca gcggtgttgc
     3841 gcctcgtggg ctgatgcggc ccagcagcat cagcccgtcg atgatgagca ccgcacccac
     3901 gtagaagagc ccgacgccac ccatgtcgct agtacctcgg cagactcggt agaccgggat
     3961 tgcggcgcgg ggcccgcggg cggtccgcgc ctgcgggaat cgacttcacc accgtgggac
     4021 tttcggcgct cgcggacgcc gcgtcgaatg cgcggtgatg tcccgcggtg aatgtcgtca
     4081 gccctaccga accgaagacc cggcgcgccg gcgtccggca ctccggacag gtcgccgccg
     4141 catcgcgctc ggagatcgcg caggtgagat cgaagggacc gcagtgggaa caacggaatg
     4201 tgtaggtcgg catggtcacc cctttccatt cacccgaaat atatccagat gaaaataaaa
     4261 gtcaagggtg aacgcccgcg ccgggcgcgg gtggttacga agtcaaattc gaggtggtgt
     4321 cgacgagttc gcgcaactgg tcgacgtcca gcccgcacga ttcaaaagcc gcacactcgg
     4381 cctcggctac cgcgtccacc aggcggttgc gcagcgtccg cccccgcggg gtcaacgcca
     4441 cgagcacccg ccggcggtca tgcagatcgg cagtccggta gatgatccct tgcgacacaa
     4501 ggcgatcgac ggttctggtg agagtcggac ccgtgatcag cgtggcctgc gcgagatcgc
     4561 tcatgggcac ggtgcgccgt ccggccaact cgtcgagcac gcgccagtcc tcgatcttga
     4621 gcgacgagtc cgcgagcacg cgttcgatcg ccgactccgc gaggtgggtc agacgcacca
     4681 acggcagcag gaggttgggg cggcgctgct gtccggcggc catggtgggt gtctcctcgt
     4741 gtcgacggcg gggtctgttc agaatgaatt atttctgcta gaaagctttg aagcgcagac
     4801 tacacgggat cgacggtgat ctggagtgca ggacggcgag gttgagttcc gcgtcgggct
     4861 ggtgattccg cttcaagggc cagcgggaat cttcgcgcct tcgtgcgagg ccgtggccga
     4921 gctcgcggcc aaggaggtca acgaccgagg cggcctgcag ggccgcaagg tcaccatcga
     4981 ggtgctcgac ggcggagcgc ccggcgacga cgtcgcccgg accgtcgccg accggttgcg
     5041 aggtcacggt ctcgacgccg tgaccggctg gcacatctcg gccgtgcgca accgcatctc
     5101 cccggtggtc cgcgaccgca tcccgtacgt ctacacctcg ttgtacgagg gcggtgaacg
     5161 cacaccgggc gtgttctgca caggcgagac accgcagatc cagatcgcgc ccgcgctcgc
     5221 ctggctgcgc gaccacttcg gcatccggtc ctggtgcctg gtcggcgatg actacatctg
     5281 gccgcgccgt tccgccgcgg ccgcccgcgc gtactgccga gatctggacc tcgagctcag
     5341 acgggagatc tacgtcccgt acggcaccga cgatttccgc gcacctgtcc gcaaggccat
     5401 cgcgtcgggg gcgcaggccg tgttgatgct gctcgtcggc caggacgccg tgttgttcaa
     5461 ccgcgagttc gcgcgcgccg gcgggcacga ccgcatggcg cggttcagcc cgctgatgga
     5521 ggagaacatg ctgctggcca gcggcgccgg ctccaccgaa aacctctatg tcgccgcggc
     5581 gtacttcagt tcactggcca ccgcgggtgc catggacctg atgggcagct acgtcgcccg
     5641 ctacggcgcc gacgcgccac cgctcaacgc gatggccgaa tcctgttacg agggtctgct
     5701 tgcgctcgag gccatcttcc agcgggccca ctcccccgag ataccggacc tgatggcatc
     5761 cgcgcacgat gtcgggttcg acgggccccg gggcccgatg tgtatgcgcg acagtcaatt
     5821 cgaccagcag gtctacatcg ccagcgccga cggctacgac ttcgacatcc tcgacacgct
     5881 ggcgacgctc gacgcttgag accgcgtcac ttctttatct agatttaaag atctggtacc
     5941 gcggccgcgc cacggatgcc accacaggca ctacaacgga gttcgccacg tacatcacca
     6001 caaccaccga ttctggcggt gagctccccg atattcagcg gaaatggctt ggtatcgacc
     6061 aagattcgta gaaccccgtc tcgtctggct ggtattcaaa acgggcgcaa cgaaacacgc
     6121 aacgagacag gcatggccca aaccagaaaa ctagcgtcta ccaggacttt tacctgtccg
     6181 acccgttgca acggaacccc ccacggaacc cccgcgacac ccgctcccca attgcgttag
     6241 aacagcggtg gattgtcggc ttcgttgtgg gccttttgag ccgcttcctg ttctgccgca
     6301 cgctctttcc tcgcccgata gccgagtcgc ttaacggtgt ccagatgcag cccgaaatgt
     6361 ttggccgttt gcggccaaga gtggccctcg tcgtcgtgat aggcgcggat gcgttcgcgg
     6421 cgtgcagcct gctcggcgag ccactcgctg cgttcctgcg ccacgagccg gacgacgtgg
     6481 cgttcggata gtccggtgat tcgagcgcct tcggcggcgg tcacgcgccg ctttttgcgg
     6541 acagtcggct gccggttgta gccgtcgctg tagccgtcgc tcatagcaat gcctccatgg
     6601 ctgacgcgga ctttgcgcgc cgcgcaactg tgctcgccgc cgtgcgcgct gctgcgccct
     6661 tccgcgagat ggccgactgg cgcgcactga gtgtggcctc gtagaccacg atcccgtccg
     6721 cccaaatgcg cgacttggtt gtgatccaac gccaaatgct gttggcgatg gcgcggacct
     6781 cgctgtccgg tagcggtccg ggacacacgt cgttgcacgg aaattcggcg tttcgcgcgt
     6841 ggcactcggc atagatcgcg cggccgagtc cgtccacgtt ccgggtcggc aggtagatcc
     6901 gcatgagggc gggacgatag gcccacaacc tgacggaatc gaacagtgcg caattccgcc
     6961 ctagcggcgt cggagccgct ttgtacgtgg tctgctgacg ccagcgcggc ggtggcatgt
     7021 tcgcgccgag ctcggcctcg atgtggctga gtgtgtagag atctgagtgg agccattccg
     7081 tttcccaggc gatgtggccg gggtttttgg tcatgaggcc tgagtaactg cggtcgccat
     7141 cgacggcgcg ccgaaggcct tcggcgcacg ccgccatgta tgcgagcggc ttacgccgcg
     7201 cgtattcggt gcgtggaaca ggggcgttga gtgcccacac tgcgtgtgcg tggccgttgg
     7261 cgcgattgcc cacgatcgcg ttgggcagcg gatgggaccc ccgggcgctg agcgctcgga
     7321 gcgctgcgtc tggatggtct acgtccacga ccagcaggtt tgccagcgct gttgggttcg
     7381 cctcgatgta ccggcggcct agggccgacg cgcggctttg gcggtagatc ccctcgagca
     7441 gatcgtcgct tgccagcggc cagtacggca gccagagctg ctcaaattcg tcggcgacgt
     7501 ggctcacgct tggtagtaga ccacgattaa tcaccggtgt atggtccgac acgagctcca
     7561 agtcagatat ttcgctgagg ggccacccca caactgcaca ctcccccgct ctcccgtcga
     7621 gccctggtgg tggaacacca gcgacagccg agcaccccca accacctgta ccaaccagga
     7681 ggaacacatg cgtcgtttcg aggacgtttc cgggccgctg agagccgctg tggcggccgt
     7741 acacgccgcc ttagacccgt tagaccccct gccgcctgaa tgcgcgggta cgagccacac
     7801 agcgcccgaa cttacggagc tggtgggctc acgcgttgcg ctcggtcgtt cggctgcggc
     7861 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7921 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7981 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8041 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8101 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8161 cctcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8221 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8281 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8341 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8401 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     8461 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8521 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8581 aagatccttt gatcttttct acggggtctg acgctcatgg aactagtgag gtctgcctcg
     8641 tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt
     8701 gagggagcca cggttggtga gagctttgtt gtaggtggac cagttggtga ttttgaactt
     8761 ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc
     8821 agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc
     8881 cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     8941 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     9001 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     9061 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     9121 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagattatgc
     9181 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     9241 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     9301 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     9361 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     9421 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     9481 ggaagaggca taaattccgt cagcc
//
