LOCUS       sequence-482711-        9264 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  pLAM12 vector backbone containing Eas1 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 9264)
  TITLE     Shipman lab plasmids 2025
REFERENCE   2  (bases 1 to 9264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRECMYC-Eas1
FEATURES             Location/Qualifiers
     source          1..9264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      6380..6426
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6576..6595)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6587..7402
                     /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(6660..6679)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7733..8321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8222..8241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccacatcga ggccgagctc ggcgcgaaca tgccaccgcc gcgctggcgt cagcagacca
       61 cgtacaaagc ggctccgacg ccgctagggc ggaattgcgc actgttcgat tccgtcaggt
      121 tgtgggccta tcgtcccgcc ctcatgcgga tctacctgcc gacccggaac gtggacggac
      181 tcggccgcgc gatctatgcc gagtgccacg cgcgaaacgc cgaatttccg tgcaacgacg
      241 tgtgtcccgg accgctaccg gacagcgagg tccgcgccat cgccaacagc atttggcgtt
      301 ggatcacaac caagtcgcgc atttgggcgg acgggatcgt ggtctacgag gccacactca
      361 gtgcgcgcca gtcggccatc tcgcggaagg gcgcagcagc gcgcacggcg gcgagcacag
      421 ttgcgcggcg cgcaaagtcc gcgtcagcca tggaggcatt gctatgagcg acggctacag
      481 cgacggctac aaccggcagc cgactgtccg caaaaagcgg cgcgtgaccg ccgccgaagg
      541 cgctcgaatc accggactat ccgaacgcca cgtcgtccgg ctcgtggcgc aggaacgcag
      601 cgagtggctc gccgagcagg ctgcacgccg cgaacgcatc cgcgcctatc acgacgacga
      661 gggccactct tggccgcaaa cggccaaaca tttcgggctg catctggaca ccgttaagcg
      721 actcggctat cgggcgagga aagagcgtgc ggcagaacag gaagcggctc aaaaggccca
      781 caacgaagcc gacaatccac cgctgttcta acgcaattgg ggagcgggtg tcgcgggggt
      841 tccgtggggg gttccgttgc aacgggtcgg acaggtaaaa gtcctggtag acgctagttt
      901 tctggtttgg gccatgcctg tctcgttgcg tgtttcgttg cgcccgtttt gaataccagc
      961 cagacgagac ggggttctac gaatcttggt cgataccaag ccatttccgc tgaatatcgg
     1021 ggagctcacc gccagaatcg gtggttgtgg tgatgtacgt ggcgaactcc gttgtagtgc
     1081 ctgtggtggc atccgtggcg cggccgcggt accagatctt taaatctaga taaagaagtg
     1141 acgcggtctc aagcgtcgag cgtcgccagc gtgtcgagga tgtcgaagtc gtagccgtcg
     1201 gcgctggcga tgtagacctg ctggtcgaat tgactgtcgc gcatacacat cgggccccgg
     1261 ggcccgtcga acccgacatc gtgcgcggat gccatcaggt ccggtatctc gggggagtgg
     1321 gcccgctgga agatggcctc gagcgcaagc agaccctcgt aacaggattc ggccatcgcg
     1381 ttgagcggtg gcgcgtcggc gccgtagcgg gcgacgtagc tgcccatcag gtccatggca
     1441 cccgcggtgg ccagtgaact gaagtacgcc gcggcgacat agaggttttc ggtggagccg
     1501 gcgccgctgg ccagcagcat gttctcctcc atcagcgggc tgaaccgcgc catgcggtcg
     1561 tgcccgccgg cgcgcgcgaa ctcgcggttg aacaacacgg cgtcctggcc gacgagcagc
     1621 atcaacacgg cctgcgcccc cgacgcgatg gccttgcgga caggtgcgcg gaaatcgtcg
     1681 gtgccgtacg ggacgtagat ctcccgtctg agctcgaggt ccagatctcg gcagtacgcg
     1741 cgggcggccg cggcggaacg gcgcggccag atgtagtcat cgccgaccag gcaccaggac
     1801 cggatgccga agtggtcgcg cagccaggcg agcgcgggcg cgatctggat ctgcggtgtc
     1861 tcgcctgtgc agaacacgcc cggtgtgcgt tcaccgccct cgtacaacga ggtgtagacg
     1921 tacgggatgc ggtcgcggac caccggggag atgcggttgc gcacggccga gatgtgccag
     1981 ccggtcacgg cgtcgagacc gtgacctcgc aaccggtcgg cgacggtccg ggcgacgtcg
     2041 tcgccgggcg ctccgccgtc gagcacctcg atggtgacct tgcggccctg caggccgcct
     2101 cggtcgttga cctccttggc cgcgagctcg gccacggcct cgcacgaagg cgcgaagatt
     2161 cccgctggcc cttgaagcgg aatcaccagc ccgacgcgga actcaacctc gccgtcctgc
     2221 actccagatc accgtcgatc ccgtgtagtc tgcgcttcaa agctttctag cagaaataat
     2281 tcattctgaa cagaccccgc cgtcgacacg aggagacacc caccatggcc gccggacagc
     2341 agcgccgccc caacctcctg ctgccgttgg tgcgtctgac ccacctcgcg gagtcggcga
     2401 tcgaacgcgt gctcgcggac tcgtcgctca agatcgagga ctggcgcgtg ctcgacgagt
     2461 tggccggacg gcgcaccgtg cccatgagcg atctcgcgca ggccacgctg atcacgggtc
     2521 cgactctcac cagaaccgtc gatcgccttg tgtcgcaagg gatcatctac cggactgccg
     2581 atctgcatga ccgccggcgg gtgctcgtgg cgttgacccc gcgggggcgg acgctgcgca
     2641 accgcctggt ggacgcggta gccgaggccg agtgtgcggc ttttgaatcg tgcgggctgg
     2701 acgtcgacca gttgcgcgaa ctcgtcgaca ccacctcgaa tttgacttcg taaccacccg
     2761 cgcccggcgc gggcgttcac ccttgacttt tattttcatc tggatatatt tcgggtgaat
     2821 ggaaaggggt gaccatgccg acctacacat tccgttgttc ccactgcggt cccttcgatc
     2881 tcacctgcgc gatctccgag cgcgatgcgg cggcgacctg tccggagtgc cggacgccgg
     2941 cgcgccgggt cttcggttcg gtagggctga cgacattcac cgcgggacat caccgcgcat
     3001 tcgacgcggc gtccgcgagc gccgaaagtc ccacggtggt gaagtcgatt cccgcaggcg
     3061 cggaccgccc gcgggccccg cgccgcaatc ccggtctacc gagtctgccg aggtactagc
     3121 gacatgggtg gcgtcgggct cttctacgtg ggtgcggtgc tcatcatcga cgggctgatg
     3181 ctgctgggcc gcatcagccc acgaggcgca acaccgctga acttcttcgt cggcggactg
     3241 caggtggtga cgcctacggt gctgatcctg cagtccggcg gagacgcggc cgtgatcttc
     3301 gcggcctccg ggctctacct gttcggcttc acctacctgt gggtggccat caacaacgtg
     3361 accgactggg acggagaagg tctcggatgg ttctcgctgt tcgtcgcgat cgccgcactc
     3421 ggctactcgt ggcacgcgtt caccgccgag gccgacccgg cgttcggggt gatctggctg
     3481 ctgtgggcag tgctgtggtt catgctgttc ctgctgctcg gcctggggca cgacgcactg
     3541 gggcccgccg tcgggttcgt cgcggtggcc gaaggcgtga tcaccgccgc cgtgccggcc
     3601 ttcctgatcg tgtcgggcaa ctgggaaacc ggcccgctcc ccgccgcggt catcgccgtg
     3661 atcggttttg ccgcagttgt tctcgcatac cccatcgggc gccgtctcgc agcgccgtca
     3721 gtcaccaacc ctccaccggc cgcgctcgcg gccaccaccc gataagagaa agggagtcca
     3781 catatgaccc aatggaatcc gaatctgttc cgtaaggaag gtctcagcaa gggttacgat
     3841 agccactatc tcacccgttt ggtggagcaa ggcgaaacca tcaacgagcg caatctgcca
     3901 gtcgtgtttt cgttggccca cttggccaac ttgtcccaca ccctgtatag cgatctccac
     3961 ggtttcgtca gccgtaacga tctggagcgg aaggatttcc cctacaagaa cttttcgatt
     4021 agcaaacgga cgggtggcaa acggtggatt tccattccaa cgccggcgtt gatggccgtg
     4081 caatactgga tcaacaaaaa tattctcaat cggatcatcc cgcatgcggc ggcgtatgcg
     4141 tatgtgccca agcgtaagat tattcagcac gcgcaacgcc attgcggctc gggttggctc
     4201 attaaaattg atatcaaaga cttttttggt aacattagcg aacgtcaagt cttcgacgtc
     4261 tttctcaact tgggctaccc aaaactgttg agctttgaaa tggcgcgtct ctgcacccgc
     4321 gtgaccccaa aacggaaagg taagcgttgg tgtaacacca acaatgaaca cacgatcacg
     4381 ggttatgata gcaagttcgt cggttccctg ccacaaggcg cccccacgtc gccagccctg
     4441 tccaacctcg tctgtatcgg tctcgacaag cagctcacgt cgttggccat ttcggagggc
     4501 gcgacgtatt cccggtatgc ggacgacatc tgtttttccc tgccgatggg ctgtcggaaa
     4561 tccgccttgc atattaagaa ggagattagc aagattttgc acggttacgg gttttcggat
     4621 aacagcaaga aaacccgtct catcccgccg ggggcgcgta aggtgctcac cgggctgaac
     4681 gtcaataacg ataccccacg cattccaaaa gagatccgtg atagcgtgcg tatgcacctc
     4741 tactatgcga agaaaaacgg catcgcgacg cattgtaaaa cgcgcggttt ccggagcgtc
     4801 attggctttt acaattacct ctcgggggtg atcggttata tttactcgat tgattcgaat
     4861 atggggaaca agttccggaa tgagttcaac tcgttgccgt gggtcgactt ctccttgtaa
     4921 taccgttcat atgaggctag agcaggtgat tggggttccg tcgtgaaact tagttgtggt
     4981 agatacatat atagtgtatc aactgccatt ggcagcctac gattccgttt gaatggtttg
     5041 acaggatgcc aacgagcacg gattccgtcg ctctttgtag tagccttcgc cagagtgagt
     5101 gcgccgtcgt gcgtcacgcc cggcggggcc cgtgtgagcc gctggacacc atcaccccgg
     5161 cgccagaggc tcacttacgt gaggtgaagt aagtgactca atggaacccc agccacctgc
     5221 atgatcccgg cgggagaaag gagaaagcca atcatggctg aaaatgctgt caccaagcag
     5281 gattcgccca aggcacccga gacgatctcg caagtgctgc aggtgctcgt gccgcagctg
     5341 gcgcgtgcag tgcccaaggg tatggacccc gaccgcatcg cgcggatcgt gcaaaccgag
     5401 atccgcaagt cgcgcaacgc gaaagctgct ggcatcgcta agcagtccct cgacgactgc
     5461 acgcaagagt catttgccgg tgcgctgctg acctcggccg cgctcggtct cgagcccggt
     5521 gtcaacggcg agtgctacct cgtgccctac cgcgacaccc ggcgcggcgt ggtcgagtgc
     5581 cagctgatca tcggctacca gggcatcgtc aaactgttct ggcagcaccc gcgcgcctcc
     5641 cggatcgacg cgcagtgggt cggcgcgaac gacgaattcc attacaccat gggcctcaat
     5701 ccgacgctga aacacgtgaa ggccaagggt gatcggggta atccggtcta cttctacgcg
     5761 atcgtcgagg tgaccggcgc tgagccgctg tgggacgtgt tcaccgccga cgagatcagg
     5821 gaattgcgtc gcggcaaggt cggatcctcg ggcgacatca aggacccgca gcgctggatg
     5881 gagcggaaga ccgcgctcaa acaggtgctg aagctggcac cgaagacgac gcggctcgac
     5941 gcggcgatcc gcgccgacga tcgcccgggc accgacctgt cacagtcgca ggcgctcgcg
     6001 ctgccgtcga ccgtcaagcc gacggccgac tacatcgacg gcgagatcgc cgagccgcac
     6061 gaggtcgata cgccaccgaa gtcgtcgcgc gcacaacgcg cgcagcgcgc caccgcgccg
     6121 gcgcccgacg tgcagatggc caatcccgat cagctgaagc gcctcggcga gatccagaag
     6181 gccgaaaagt acaacgacgc cgattggttc aagttcctcg ccgattcggc cggcgtcaaa
     6241 gccacgcgcg ccgccgacct cacgttcgac gaggcgaagg ctgtcatcga catgttcgac
     6301 gggcccaacg catgagcgcc ccggcgaatt ccgacgcggt ggttgatctg caactagcgt
     6361 acgatcgact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     6421 ttttatgcca tcatggccgc ggtgatcagc tagccacctg acgtcggggg ggggggaaag
     6481 ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     6541 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     6601 acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     6661 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     6721 gaagccccat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     6781 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     6841 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     6901 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     6961 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     7021 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     7081 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataatct
     7141 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     7201 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     7261 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     7321 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     7381 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     7441 tacgctgact tgacgggacg gcggctttgt tgaataaatc gaacttttgc tgagttgaag
     7501 gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa
     7561 atcaccaact ggtccaccta caacaaagct ctcaccaacc gtggctccct cactttctgg
     7621 ctggatgatg gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga
     7681 cctcactagt tccatgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7741 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7801 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7861 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7921 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7981 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8041 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     8101 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgag gggagaaagg
     8161 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8221 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8281 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8341 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8401 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8461 gaacgaccga gcgcaacgcg tgagcccacc agctccgtaa gttcgggcgc tgtgtggctc
     8521 gtacccgcgc attcaggcgg cagggggtct aacgggtcta aggcggcgtg tacggccgcc
     8581 acagcggctc tcagcggccc ggaaacgtcc tcgaaacgac gcatgtgttc ctcctggttg
     8641 gtacaggtgg ttgggggtgc tcggctgtcg ctggtgttcc accaccaggg ctcgacggga
     8701 gagcggggga gtgtgcagtt gtggggtggc ccctcagcga aatatctgac ttggagctcg
     8761 tgtcggacca tacaccggtg attaatcgtg gtctactacc aagcgtgagc cacgtcgccg
     8821 acgaatttga gcagctctgg ctgccgtact ggccgctggc aagcgacgat ctgctcgagg
     8881 ggatctaccg ccaaagccgc gcgtcggccc taggccgccg gtacatcgag gcgaacccaa
     8941 cagcgctggc aaacctgctg gtcgtggacg tagaccatcc agacgcagcg ctccgagcgc
     9001 tcagcgcccg ggggtcccat ccgctgccca acgcgatcgt gggcaatcgc gccaacggcc
     9061 acgcacacgc agtgtgggca ctcaacgccc ctgttccacg caccgaatac gcgcggcgta
     9121 agccgctcgc atacatggcg gcgtgcgccg aaggccttcg gcgcgccgtc gatggcgacc
     9181 gcagttactc aggcctcatg accaaaaacc ccggccacat cgcctgggaa acggaatggc
     9241 tccactcaga tctctacaca ctca
//
