LOCUS       sequence-482749-        9304 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  pLAM12 vector backbone containing Hcl1 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 9304)
  TITLE     Shipman lab plasmids 2025
REFERENCE   2  (bases 1 to 9304)
  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-Hcl1
FEATURES             Location/Qualifiers
     source          1..9304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(4026..4614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4106..4125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4945..5760)
                     /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     5668..5687
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5752..5771
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(5921..5967)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgcggggccc gcgggcggtc cgcgcctgcg ggaatcgact tcaccaccgt gggactttcg
       61 gcgctcgcgg acgccgcgtc gaatgcgcgg tgatgtcccg cggtgaatgt cgtcagccct
      121 accgaaccga agacccggcg cgccggcgtc cggcactccg gacaggtcgc cgccgcatcg
      181 cgctcggaga tcgcgcaggt gagatcgaag ggaccgcagt gggaacaacg gaatgtgtag
      241 gtcggcatgg tcaccccttt ccattcaccc gaaatatatc cagatgaaaa taaaagtcaa
      301 gggtgaacgc ccgcgccggg cgcgggtggt tacgaagtca aattcgaggt ggtgtcgacg
      361 agttcgcgca actggtcgac gtccagcccg cacgattcaa aagccgcaca ctcggcctcg
      421 gctaccgcgt ccaccaggcg gttgcgcagc gtccgccccc gcggggtcaa cgccacgagc
      481 acccgccggc ggtcatgcag atcggcagtc cggtagatga tcccttgcga cacaaggcga
      541 tcgacggttc tggtgagagt cggacccgtg atcagcgtgg cctgcgcgag atcgctcatg
      601 ggcacggtgc gccgtccggc caactcgtcg agcacgcgcc agtcctcgat cttgagcgac
      661 gagtccgcga gcacgcgttc gatcgccgac tccgcgaggt gggtcagacg caccaacggc
      721 agcaggaggt tggggcggcg ctgctgtccg gcggccatgg tgggtgtctc ctcgtgtcga
      781 cggcggggtc tgttcagaat gaattatttc tgctagaaag ctttgaagcg cagactacac
      841 gggatcgacg gtgatctgga gtgcaggacg gcgaggttga gttccgcgtc gggctggtga
      901 ttccgcttca agggccagcg ggaatcttcg cgccttcgtg cgaggccgtg gccgagctcg
      961 cggccaagga ggtcaacgac cgaggcggcc tgcagggccg caaggtcacc atcgaggtgc
     1021 tcgacggcgg agcgcccggc gacgacgtcg cccggaccgt cgccgaccgg ttgcgaggtc
     1081 acggtctcga cgccgtgacc ggctggcaca tctcggccgt gcgcaaccgc atctccccgg
     1141 tggtccgcga ccgcatcccg tacgtctaca cctcgttgta cgagggcggt gaacgcacac
     1201 cgggcgtgtt ctgcacaggc gagacaccgc agatccagat cgcgcccgcg ctcgcctggc
     1261 tgcgcgacca cttcggcatc cggtcctggt gcctggtcgg cgatgactac atctggccgc
     1321 gccgttccgc cgcggccgcc cgcgcgtact gccgagatct ggacctcgag ctcagacggg
     1381 agatctacgt cccgtacggc accgacgatt tccgcgcacc tgtccgcaag gccatcgcgt
     1441 cgggggcgca ggccgtgttg atgctgctcg tcggccagga cgccgtgttg ttcaaccgcg
     1501 agttcgcgcg cgccggcggg cacgaccgca tggcgcggtt cagcccgctg atggaggaga
     1561 acatgctgct ggccagcggc gccggctcca ccgaaaacct ctatgtcgcc gcggcgtact
     1621 tcagttcact ggccaccgcg ggtgccatgg acctgatggg cagctacgtc gcccgctacg
     1681 gcgccgacgc gccaccgctc aacgcgatgg ccgaatcctg ttacgagggt ctgcttgcgc
     1741 tcgaggccat cttccagcgg gcccactccc ccgagatacc ggacctgatg gcatccgcgc
     1801 acgatgtcgg gttcgacggg ccccggggcc cgatgtgtat gcgcgacagt caattcgacc
     1861 agcaggtcta catcgccagc gccgacggct acgacttcga catcctcgac acgctggcga
     1921 cgctcgacgc ttgagaccgc gtcacttctt tatctagatt taaagatctg gtaccgcggc
     1981 cgcgccacgg atgccaccac aggcactaca acggagttcg ccacgtacat caccacaacc
     2041 accgattctg gcggtgagct ccccgatatt cagcggaaat ggcttggtat cgaccaagat
     2101 tcgtagaacc ccgtctcgtc tggctggtat tcaaaacggg cgcaacgaaa cacgcaacga
     2161 gacaggcatg gcccaaacca gaaaactagc gtctaccagg acttttacct gtccgacccg
     2221 ttgcaacgga accccccacg gaacccccgc gacacccgct ccccaattgc gttagaacag
     2281 cggtggattg tcggcttcgt tgtgggcctt ttgagccgct tcctgttctg ccgcacgctc
     2341 tttcctcgcc cgatagccga gtcgcttaac ggtgtccaga tgcagcccga aatgtttggc
     2401 cgtttgcggc caagagtggc cctcgtcgtc gtgataggcg cggatgcgtt cgcggcgtgc
     2461 agcctgctcg gcgagccact cgctgcgttc ctgcgccacg agccggacga cgtggcgttc
     2521 ggatagtccg gtgattcgag cgccttcggc ggcggtcacg cgccgctttt tgcggacagt
     2581 cggctgccgg ttgtagccgt cgctgtagcc gtcgctcata gcaatgcctc catggctgac
     2641 gcggactttg cgcgccgcgc aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc
     2701 gagatggccg actggcgcgc actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa
     2761 atgcgcgact tggttgtgat ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg
     2821 tccggtagcg gtccgggaca cacgtcgttg cacggaaatt cggcgtttcg cgcgtggcac
     2881 tcggcataga tcgcgcggcc gagtccgtcc acgttccggg tcggcaggta gatccgcatg
     2941 agggcgggac gataggccca caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc
     3001 ggcgtcggag ccgctttgta cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg
     3061 ccgagctcgg cctcgatgtg gctgagtgtg tagagatctg agtggagcca ttccgtttcc
     3121 caggcgatgt ggccggggtt tttggtcatg aggcctgagt aactgcggtc gccatcgacg
     3181 gcgcgccgaa ggccttcggc gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat
     3241 tcggtgcgtg gaacaggggc gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga
     3301 ttgcccacga tcgcgttggg cagcggatgg gacccccggg cgctgagcgc tcggagcgct
     3361 gcgtctggat ggtctacgtc cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg
     3421 atgtaccggc ggcctagggc cgacgcgcgg ctttggcggt agatcccctc gagcagatcg
     3481 tcgcttgcca gcggccagta cggcagccag agctgctcaa attcgtcggc gacgtggctc
     3541 acgcttggta gtagaccacg attaatcacc ggtgtatggt ccgacacgag ctccaagtca
     3601 gatatttcgc tgaggggcca ccccacaact gcacactccc ccgctctccc gtcgagccct
     3661 ggtggtggaa caccagcgac agccgagcac ccccaaccac ctgtaccaac caggaggaac
     3721 acatgcgtcg tttcgaggac gtttccgggc cgctgagagc cgctgtggcg gccgtacacg
     3781 ccgccttaga cccgttagac cccctgccgc ctgaatgcgc gggtacgagc cacacagcgc
     3841 ccgaacttac ggagctggtg ggctcacgcg ttgcgctcgg tcgttcggct gcggcgagcg
     3901 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3961 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccctcg
     4201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4441 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4561 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4621 cctttgatct tttctacggg gtctgacgct catggaacta gtgaggtctg cctcgtgaag
     4681 aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg
     4741 agccacggtt ggtgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct
     4801 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
     4861 aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     4921 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     4981 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     5041 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     5101 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     5161 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagat tatgcatttc
     5221 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     5281 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     5341 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     5401 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     5461 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     5521 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     5581 gctacctttg ccatgtttca gaaacaactc tggcgcatgg ggcttcccat acaagcgata
     5641 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     5701 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
     5761 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     5821 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt cccccccccc
     5881 cgacgtcagg tggctagctg atcaccgcgg ccatgatggc ataaaacgaa aggcccagtc
     5941 tttcgactga gcctttcgtt ttatttgatg cctggcagtc gatcgtacgc tagttgcaga
     6001 tcaaccaccg cgtcggaatt cgccggggcg ctcatgcgtt gggcccgtcg aacatgtcga
     6061 tgacagcctt cgcctcgtcg aacgtgaggt cggcggcgcg cgtggctttg acgccggccg
     6121 aatcggcgag gaacttgaac caatcggcgt cgttgtactt ttcggccttc tggatctcgc
     6181 cgaggcgctt cagctgatcg ggattggcca tctgcacgtc gggcgccggc gcggtggcgc
     6241 gctgcgcgcg ttgtgcgcgc gacgacttcg gtggcgtatc gacctcgtgc ggctcggcga
     6301 tctcgccgtc gatgtagtcg gccgtcggct tgacggtcga cggcagcgcg agcgcctgcg
     6361 actgtgacag gtcggtgccc gggcgatcgt cggcgcggat cgccgcgtcg agccgcgtcg
     6421 tcttcggtgc cagcttcagc acctgtttga gcgcggtctt ccgctccatc cagcgctgcg
     6481 ggtccttgat gtcgcccgag gatccgacct tgccgcgacg caattccctg atctcgtcgg
     6541 cggtgaacac gtcccacagc ggctcagcgc cggtcacctc gacgatcgcg tagaagtaga
     6601 ccggattacc ccgatcaccc ttggccttca cgtgtttcag cgtcggattg aggcccatgg
     6661 tgtaatggaa ttcgtcgttc gcgccgaccc actgcgcgtc gatccgggag gcgcgcgggt
     6721 gctgccagaa cagtttgacg atgccctggt agccgatgat cagctggcac tcgaccacgc
     6781 cgcgccgggt gtcgcggtag ggcacgaggt agcactcgcc gttgacaccg ggctcgagac
     6841 cgagcgcggc cgaggtcagc agcgcaccgg caaatgactc ttgcgtgcag tcgtcgaggg
     6901 actgcttagc gatgccagca gctttcgcgt tgcgcgactt gcggatctcg gtttgcacga
     6961 tccgcgcgat gcggtcgggg tccataccct tgggcactgc acgcgccagc tgcggcacga
     7021 gcacctgcag cacttgcgag atcgtctcgg gtgccttggg cgaatcctgc ttggtgacag
     7081 cattttcagc catgattggc tttctccttt ctcccgccgg gatcatgcag gtggcatcat
     7141 tgacaacacc gtcctcctta cctttattcc ttttgatgaa gtaagcctct ggcgccgggg
     7201 tgatggtgtc cagcggctca cacgggcccc gccgggcgtg acgcacgacg gcgcacttac
     7261 ttcattaaat atagtcctga catagcacct cggaaagcga accacttccc actactttca
     7321 tataaataaa tattgtgcac gtttatacaa aagtagtagc atctaaggat aacgatgttg
     7381 tcaataatat cacctgctct agcctcatat gaacggtact actcgtacgt ttctttcgtg
     7441 gcggccgcgt tcgtcttgaa ttcatcttgc tgcgactgaa tgtgcgcgga caaaaagtcc
     7501 actttcccga ttttaccgag atactcttgc cgcgcggaca caattttctc cagctccaag
     7561 gacacttgat cgtcatagtc ataaccctta tagttcatat cctggatctc tttgaaaatt
     7621 ttgttatcat taaagcgttc cacgatttcc tcgaaaaacg cgtagtcttt attcgagagc
     7681 gagttaatgt aggtgatcac tttttcgcga taacccactt caatcgaatt gatatacgag
     7741 acgatgccgc gaatgcgatc aatttgggag taatcggccg tgacaatggc ttgatggatc
     7801 atggcacgga cggtacgttt caactgtttg ctcgctttga ttttggaatc gttgaccgta
     7861 ataccggtga cttttttgtg cgagccgggg gacaggagac gcgtcttgtt cttgttgacc
     7921 ttgaaacctt cgtcttcaat gatattaata ataatgttaa tggccttcgt gagcgtctgc
     7981 ttattatcgg acgaaaaggt caaatcatcc gcatagcggg tatagagaat atcccgcttg
     8041 ccacacaagc ccgacaggcg gttgtcgaga cgataacaga tcaaattcga aatgtatggg
     8101 gaacacacac ccccctgcgg cagataacca ttataggtac acaaattggt cagaatattg
     8161 ctcaccatgt tgttatagcc caggttcttg aacaagtaaa agacccgttc acgcttgatg
     8221 gaatgaaaaa aatctttgat gtccatttcc agaatataca agctataccg atggtgcatg
     8281 gcattttttt tgatcccgtt atttttccct ttcttatagg ccatggcctg ttcggacaag
     8341 ctcactttct cgagaatctc tttcaagatc caccgctgaa tcaatttgag ggaatacgtc
     8401 ggggacaaga tctcacggct cgtgccgtcg cgtttcttga tataaaaacg tttgtagtag
     8461 ttttccgtct tcttggtcag caagaagatc aggcgtttgg aaatcgcgaa cgcgtccgag
     8521 aggctgtcca gatcgctaat ggtgggcaag ttcagcgcat tcagaatgaa gctgtcggtg
     8581 taatccttct ggatcatgct caaatgtgga ctccctttct cttatcgggt ggtggccgcg
     8641 agcgcggccg gtggagggtt ggtgactgac ggcgctgcga gacggcgccc gatggggtat
     8701 gcgagaacaa ctgcggcaaa accgatcacg gcgatgaccg cggcggggag cgggccggtt
     8761 tcccagttgc ccgacacgat caggaaggcc ggcacggcgg cggtgatcac gccttcggcc
     8821 accgcgacga acccgacggc gggccccagt gcgtcgtgcc ccaggccgag cagcaggaac
     8881 agcatgaacc acagcactgc ccacagcagc cagatcaccc cgaacgccgg gtcggcctcg
     8941 gcggtgaacg cgtgccacga gtagccgagt gcggcgatcg cgacgaacag cgagaaccat
     9001 ccgagacctt ctccgtccca gtcggtcacg ttgttgatgg ccacccacag gtaggtgaag
     9061 ccgaacaggt agagcccgga ggccgcgaag atcacggccg cgtctccgcc ggactgcagg
     9121 atcagcaccg taggcgtcac cacctgcagt ccgccgacga agaagttcag cggtgttgcg
     9181 cctcgtgggc tgatgcggcc cagcagcatc agcccgtcga tgatgagcac cgcacccacg
     9241 tagaagagcc cgacgccacc catgtcgcta gtacctcggc agactcggta gaccgggatt
     9301 gcgg
//
