LOCUS       sequence-483558-        9799 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  pLAM12 vector backbone containing Eco2 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 9799)
  TITLE     Shipman lab plasmids 2025
REFERENCE   2  (bases 1 to 9799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRECMYC-Eco2
FEATURES             Location/Qualifiers
     source          1..9799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3046..3634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3126..3145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3965..4780)
                     /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     4688..4707
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4772..4791
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(4941..4987)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgaggcggcc tgcagggccg caaggtcacc atcgaggtgc tcgacggcgg agcgcccggc
       61 gacgacgtcg cccggaccgt cgccgaccgg ttgcgaggtc acggtctcga cgccgtgacc
      121 ggctggcaca tctcggccgt gcgcaaccgc atctccccgg tggtccgcga ccgcatcccg
      181 tacgtctaca cctcgttgta cgagggcggt gaacgcacac cgggcgtgtt ctgcacaggc
      241 gagacaccgc agatccagat cgcgcccgcg ctcgcctggc tgcgcgacca cttcggcatc
      301 cggtcctggt gcctggtcgg cgatgactac atctggccgc gccgttccgc cgcggccgcc
      361 cgcgcgtact gccgagatct ggacctcgag ctcagacggg agatctacgt cccgtacggc
      421 accgacgatt tccgcgcacc tgtccgcaag gccatcgcgt cgggggcgca ggccgtgttg
      481 atgctgctcg tcggccagga cgccgtgttg ttcaaccgcg agttcgcgcg cgccggcggg
      541 cacgaccgca tggcgcggtt cagcccgctg atggaggaga acatgctgct ggccagcggc
      601 gccggctcca ccgaaaacct ctatgtcgcc gcggcgtact tcagttcact ggccaccgcg
      661 ggtgccatgg acctgatggg cagctacgtc gcccgctacg gcgccgacgc gccaccgctc
      721 aacgcgatgg ccgaatcctg ttacgagggt ctgcttgcgc tcgaggccat cttccagcgg
      781 gcccactccc ccgagatacc ggacctgatg gcatccgcgc acgatgtcgg gttcgacggg
      841 ccccggggcc cgatgtgtat gcgcgacagt caattcgacc agcaggtcta catcgccagc
      901 gccgacggct acgacttcga catcctcgac acgctggcga cgctcgacgc ttgagaccgc
      961 gtcacttctt tatctagatt taaagatctg gtaccgcggc cgcgccacgg atgccaccac
     1021 aggcactaca acggagttcg ccacgtacat caccacaacc accgattctg gcggtgagct
     1081 ccccgatatt cagcggaaat ggcttggtat cgaccaagat tcgtagaacc ccgtctcgtc
     1141 tggctggtat tcaaaacggg cgcaacgaaa cacgcaacga gacaggcatg gcccaaacca
     1201 gaaaactagc gtctaccagg acttttacct gtccgacccg ttgcaacgga accccccacg
     1261 gaacccccgc gacacccgct ccccaattgc gttagaacag cggtggattg tcggcttcgt
     1321 tgtgggcctt ttgagccgct tcctgttctg ccgcacgctc tttcctcgcc cgatagccga
     1381 gtcgcttaac ggtgtccaga tgcagcccga aatgtttggc cgtttgcggc caagagtggc
     1441 cctcgtcgtc gtgataggcg cggatgcgtt cgcggcgtgc agcctgctcg gcgagccact
     1501 cgctgcgttc ctgcgccacg agccggacga cgtggcgttc ggatagtccg gtgattcgag
     1561 cgccttcggc ggcggtcacg cgccgctttt tgcggacagt cggctgccgg ttgtagccgt
     1621 cgctgtagcc gtcgctcata gcaatgcctc catggctgac gcggactttg cgcgccgcgc
     1681 aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc gagatggccg actggcgcgc
     1741 actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa atgcgcgact tggttgtgat
     1801 ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg tccggtagcg gtccgggaca
     1861 cacgtcgttg cacggaaatt cggcgtttcg cgcgtggcac tcggcataga tcgcgcggcc
     1921 gagtccgtcc acgttccggg tcggcaggta gatccgcatg agggcgggac gataggccca
     1981 caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc ggcgtcggag ccgctttgta
     2041 cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg ccgagctcgg cctcgatgtg
     2101 gctgagtgtg tagagatctg agtggagcca ttccgtttcc caggcgatgt ggccggggtt
     2161 tttggtcatg aggcctgagt aactgcggtc gccatcgacg gcgcgccgaa ggccttcggc
     2221 gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat tcggtgcgtg gaacaggggc
     2281 gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga ttgcccacga tcgcgttggg
     2341 cagcggatgg gacccccggg cgctgagcgc tcggagcgct gcgtctggat ggtctacgtc
     2401 cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg atgtaccggc ggcctagggc
     2461 cgacgcgcgg ctttggcggt agatcccctc gagcagatcg tcgcttgcca gcggccagta
     2521 cggcagccag agctgctcaa attcgtcggc gacgtggctc acgcttggta gtagaccacg
     2581 attaatcacc ggtgtatggt ccgacacgag ctccaagtca gatatttcgc tgaggggcca
     2641 ccccacaact gcacactccc ccgctctccc gtcgagccct ggtggtggaa caccagcgac
     2701 agccgagcac ccccaaccac ctgtaccaac caggaggaac acatgcgtcg tttcgaggac
     2761 gtttccgggc cgctgagagc cgctgtggcg gccgtacacg ccgccttaga cccgttagac
     2821 cccctgccgc ctgaatgcgc gggtacgagc cacacagcgc ccgaacttac ggagctggtg
     2881 ggctcacgcg ttgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3061 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3181 cctgccgctt accggatacc tgtccgcctt tctcccctcg ggaagcgtgg cgctttctca
     3241 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3481 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3661 gtctgacgct catggaacta gtgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     3721 caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt ggtgagagct
     3781 ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg
     3841 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     3901 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     3961 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     4021 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4081 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4141 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4201 cgactgaatc cggtgagaat ggcaaaagat tatgcatttc tttccagact tgttcaacag
     4261 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4321 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4381 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4441 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     4501 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4561 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4621 gaaacaactc tggcgcatgg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     4681 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4741 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4801 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     4861 atcagagatt ttgagacaca acgtggcttt cccccccccc cgacgtcagg tggctagctg
     4921 atcaccgcgg ccatgatggc ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     4981 ttatttgatg cctggcagtc gatcgtacgc tagttgcaga tcaaccaccg cgtcggaatt
     5041 cgccggggcg ctcatgcgtt gggcccgtcg aacatgtcga tgacagcctt cgcctcgtcg
     5101 aacgtgaggt cggcggcgcg cgtggctttg acgccggccg aatcggcgag gaacttgaac
     5161 caatcggcgt cgttgtactt ttcggccttc tggatctcgc cgaggcgctt cagctgatcg
     5221 ggattggcca tctgcacgtc gggcgccggc gcggtggcgc gctgcgcgcg ttgtgcgcgc
     5281 gacgacttcg gtggcgtatc gacctcgtgc ggctcggcga tctcgccgtc gatgtagtcg
     5341 gccgtcggct tgacggtcga cggcagcgcg agcgcctgcg actgtgacag gtcggtgccc
     5401 gggcgatcgt cggcgcggat cgccgcgtcg agccgcgtcg tcttcggtgc cagcttcagc
     5461 acctgtttga gcgcggtctt ccgctccatc cagcgctgcg ggtccttgat gtcgcccgag
     5521 gatccgacct tgccgcgacg caattccctg atctcgtcgg cggtgaacac gtcccacagc
     5581 ggctcagcgc cggtcacctc gacgatcgcg tagaagtaga ccggattacc ccgatcaccc
     5641 ttggccttca cgtgtttcag cgtcggattg aggcccatgg tgtaatggaa ttcgtcgttc
     5701 gcgccgaccc actgcgcgtc gatccgggag gcgcgcgggt gctgccagaa cagtttgacg
     5761 atgccctggt agccgatgat cagctggcac tcgaccacgc cgcgccgggt gtcgcggtag
     5821 ggcacgaggt agcactcgcc gttgacaccg ggctcgagac cgagcgcggc cgaggtcagc
     5881 agcgcaccgg caaatgactc ttgcgtgcag tcgtcgaggg actgcttagc gatgccagca
     5941 gctttcgcgt tgcgcgactt gcggatctcg gtttgcacga tccgcgcgat gcggtcgggg
     6001 tccataccct tgggcactgc acgcgccagc tgcggcacga gcacctgcag cacttgcgag
     6061 atcgtctcgg gtgccttggg cgaatcctgc ttggtgacag cattttcagc catgattggc
     6121 tttctccttt ctcccgccgg gatcatgcag gtggcacgca tgcaggcact ccttcgcctc
     6181 tggcgccggg gtgatggtgt ccagcggctc acacgggccc cgccgggcgt gacgcacgac
     6241 ggcgcacaag gattttaggg aggcgattcc tcctgccatt aaggccactg gttgcgattc
     6301 acaaccaaca aatctgccta catgcgtatc acctgctcta gcctcatatg aacggtatta
     6361 gctgttcaac atcagtttgt agtgctcctt aatatcttta atggcgtcaa agatgcagca
     6421 gaaggcttcg aaatcaattt tccgcttctt atcgcgcacg acacgcatgc taaaaatatg
     6481 tttaccgtac tccgttttcg aatcgccgtc gttgttcttg ttaaattttt tcccgtcaat
     6541 cttgatatcg agaatatctt ttgggaagag atcctccatg ctcgtttgtt cacccgatgg
     6601 gctgagtggc gtcagcacaa tatagaggtt gtagaagacg tggatatact tcattttgcg
     6661 catctcggtg acgtcatctg ggcagctttt gaccttgttg cgcaggaaat tcaggaggtc
     6721 ggaggggccc gtatcattat ccaagaccat aatgacgggc tgcttgggga cgctgccata
     6781 ataggaggcg tagttgtttt tgtaacgttc gacgaatttc ttcagatccg cggtaccacc
     6841 gctgagatcg agaaaatact tggttttttc attcgacttg aaaatgttca agttaatctc
     6901 cttcttcttg ctatcggtct tttcgcgaaa caattctgga tagctcgttt cgagcgaatg
     6961 cagggccgcc ttcaagtaaa tgcgatcggt tttaccctcg gtgatgatgg ttgggcacgt
     7021 attgccgtgg aagaacttat aataaataaa cttcgaatag gccttttcgc gcgcgttcag
     7081 cttcagttta aagccatgca gggtcgcatt ggtcaggaca tagcggtctg gttgtttatt
     7141 caatttcttt ttaatattgt taaatttatc gacttgatcg ataaacccaa acatgccttc
     7201 cagcttatcg agaccgccgg agaccaagac gccgttctcg tcgggcacct tgtattcgcc
     7261 cgtccgatac agcgcatgcg cgagcgcacg ggtctttttg taatagcagc ggtcgatatt
     7321 cacaatacgg ttgaccgtca aaccggtgac ttcttgccgc gaggtcttgt acgtcagccg
     7381 ggtcttgcta tcgttgattt cgaacccgga attttcgatc tccttgacca acaccttccc
     7441 caggacgacg ccctcgggct ggacggtcgc catttccagc gggaacgtat tcttattggt
     7501 ggaaatggtg atatcatcgg catagcggct ataggtacaa ccatactttt tggcgagttt
     7561 ggccaaacgc atatccataa tgttgcagat caagttgctg atgattggcg aacacggcga
     7621 gccttggggc agggtcccgt tatagcacgc ggccttggcc agggtggtcg ccacgacggg
     7681 attcaggaga aaatcctggt tcgacagaaa gtacccgcgc acacgcccaa aattgaagct
     7741 ttcgaagaag tccttcaagt caatgttgag aatgatctgc ttgccgcgat gcttatacgc
     7801 gttcaggatg atggacttac cacgctcaaa cccgaagctg taattgttcg aaattttacg
     7861 aatcgcaaaa atctcgtcac gacagtccga gagcaaatcg caaatacgcc gttgaatgtc
     7921 cttcagccgg tcggttgggg ccgagatggt gcggacccct ttccctttct ttggaatcgt
     7981 gaattgcgta tattggttat cggagccgat ccggtacagg acattcgtca aaaacaccag
     8041 tttgatatcg agaattttcg cgaggtcttc acgggaggtc gccgcacgga gggcatccaa
     8101 tttcgaggtt ttggtcatat gtggactccc tttctcttat cgggtggtgg ccgcgagcgc
     8161 ggccggtgga gggttggtga ctgacggcgc tgcgagacgg cgcccgatgg ggtatgcgag
     8221 aacaactgcg gcaaaaccga tcacggcgat gaccgcggcg gggagcgggc cggtttccca
     8281 gttgcccgac acgatcagga aggccggcac ggcggcggtg atcacgcctt cggccaccgc
     8341 gacgaacccg acggcgggcc ccagtgcgtc gtgccccagg ccgagcagca ggaacagcat
     8401 gaaccacagc actgcccaca gcagccagat caccccgaac gccgggtcgg cctcggcggt
     8461 gaacgcgtgc cacgagtagc cgagtgcggc gatcgcgacg aacagcgaga accatccgag
     8521 accttctccg tcccagtcgg tcacgttgtt gatggccacc cacaggtagg tgaagccgaa
     8581 caggtagagc ccggaggccg cgaagatcac ggccgcgtct ccgccggact gcaggatcag
     8641 caccgtaggc gtcaccacct gcagtccgcc gacgaagaag ttcagcggtg ttgcgcctcg
     8701 tgggctgatg cggcccagca gcatcagccc gtcgatgatg agcaccgcac ccacgtagaa
     8761 gagcccgacg ccacccatgt cgctagtacc tcggcagact cggtagaccg ggattgcggc
     8821 gcggggcccg cgggcggtcc gcgcctgcgg gaatcgactt caccaccgtg ggactttcgg
     8881 cgctcgcgga cgccgcgtcg aatgcgcggt gatgtcccgc ggtgaatgtc gtcagcccta
     8941 ccgaaccgaa gacccggcgc gccggcgtcc ggcactccgg acaggtcgcc gccgcatcgc
     9001 gctcggagat cgcgcaggtg agatcgaagg gaccgcagtg ggaacaacgg aatgtgtagg
     9061 tcggcatggt cacccctttc cattcacccg aaatatatcc agatgaaaat aaaagtcaag
     9121 ggtgaacgcc cgcgccgggc gcgggtggtt acgaagtcaa attcgaggtg gtgtcgacga
     9181 gttcgcgcaa ctggtcgacg tccagcccgc acgattcaaa agccgcacac tcggcctcgg
     9241 ctaccgcgtc caccaggcgg ttgcgcagcg tccgcccccg cggggtcaac gccacgagca
     9301 cccgccggcg gtcatgcaga tcggcagtcc ggtagatgat cccttgcgac acaaggcgat
     9361 cgacggttct ggtgagagtc ggacccgtga tcagcgtggc ctgcgcgaga tcgctcatgg
     9421 gcacggtgcg ccgtccggcc aactcgtcga gcacgcgcca gtcctcgatc ttgagcgacg
     9481 agtccgcgag cacgcgttcg atcgccgact ccgcgaggtg ggtcagacgc accaacggca
     9541 gcaggaggtt ggggcggcgc tgctgtccgg cggccatggt gggtgtctcc tcgtgtcgac
     9601 ggcggggtct gttcagaatg aattatttct gctagaaagc tttgaagcgc agactacacg
     9661 ggatcgacgg tgatctggag tgcaggacgg cgaggttgag ttccgcgtcg ggctggtgat
     9721 tccgcttcaa gggccagcgg gaatcttcgc gccttcgtgc gaggccgtgg ccgagctcgc
     9781 ggccaaggag gtcaacgac
//
