LOCUS       sequence-36465-e        4642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4642)
  AUTHORS   Stephan J, Stemmer V, Niederweis M
  TITLE     Consecutive gene deletions in Mycobacterium smegmatis using the 
            yeast FLP recombinase.
  JOURNAL   Gene. 2004 Dec 8;343(1):181-90.
  PUBMED    15563844
REFERENCE   2  (bases 1 to 4642)
  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: "Gene."; 
            date: "2004-12-8"; volume: "343(1)"; pages: "181-90"
COMMENT     Sequence Label: pMN252
FEATURES             Location/Qualifiers
     source          1..4642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1279..1312)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1366..1383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1537..2125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1617..1636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2296..3087,3088..3156))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2919..2938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3157..3261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3375..3394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    complement(4459..4492)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ggccgccggg ggccgggggc ggcgccgggc ggcccggggc gtcaggcgcc gggggcggtg
       61 tccggcggcc cccagaggaa ctgcgccagt tcctccggat cggtgaagcc ggagagatcc
      121 agcggggtct cctcgaacac ctcgaagtcg tgcaggaagg tgaaggcgag cagttcgcgg
      181 gcgaagtcct cggtccgctt ccactgcgcc ccgtcgagca gcgcggccag gatctcgcgg
      241 tcgccccgga aggcgttgag atgcagttgc accaggctgt agcgggagtc tcccgcatag
      301 acgtcggtga agtcgacgat cccggtgacc tcggtcgcgg ccaggtccac gaagatgttg
      361 gtcccgtgca ggtcgccgtg gacgaaccgg ggttcgcggc cggccagcag cgtgtccacg
      421 tccggcagcc agtcctccag gcggtccagc agccggggcg agaggtagcc ccacccgcgg
      481 tggtcctcga cggtcgccgc gcggcgttcc cgcagcagtt ccgggaagac ctcggaatgg
      541 ggggtgagca cggtgttccc ggtcagcggc accctgtgca gccggccgag cacccggccg
      601 agttcgcggg ccagggcgag cagcgcgttc cggtcggtcg tgccgtccat cgcggaccgc
      661 caggtggtgc cggtcatccg gctcatcacc aggtagggcc acggccaggc tccggtgccg
      721 ggccgcagct cgccgcggcc gaggaggcgg ggcaccggca ccggggcgtc cgccaggacc
      781 gcgtacgcct ccgactccga cgcgaggctc tccggaccgc accagtgctc gccgaacagc
      841 ttgatcaccg ggtcgggctc gccgaccagt acggggttgg tgctctcgcc gggcacccgc
      901 agcaccggcg gcaccggcag cccgagctcc tccagggctc ggcgggccag cggctcccag
      961 aattcctggt cgttccgcag gctcgcgtag gaatcatccg aatcaatacg gtcgagaagt
     1021 aacagggatt cttgtgtcac agcggacctc tattcacagg gtacgggccg gcttaattcc
     1081 gcacggccgg tcgcgacacg gcctgtccgc accgcggtca ggcgttgacg atgacgggct
     1141 ggtcggccac gtccggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1201 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1261 tagattgatt tcgttaacga agttcctata ctttctagag aataggaact tcactagtac
     1321 ctggttcgcg agtttaaacg ggagcttatc gataccgtcg gcttcctcgc tcactgactc
     1381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccctagt
     3421 ggatctaaaa aaaagcccgc tcattaggcg ggctagatcg ggtcggtact cggcgttcac
     3481 ccggatgccg ctgggcggtg acgatcccga atacaccgct gcgactaggg gcgcagccgc
     3541 gcccgtcatc gccgtgctgt cctcgtacgg cctcgacggt gagcaggctt tctacgcggc
     3601 gctcgagttt tggtcggcac tgcatgggtt tctgttgctg gaaacgaccg gcgtcatgga
     3661 cgacatcgat accgatgcgg tgttcaccga catggtgctg cggctggcgg cgggcatgga
     3721 aaggcgcacc acacacggtg gtaccgcgtc aacgtagcgc cctgcttcgg ccgcaacgcc
     3781 cgctttgacc tgccagactg gcggcgggta ttgtggttgc tcgtgcctgg cggcttacgc
     3841 ttgatgtagg ggcgtttgat gccgggccaa ttcgcatgtc cgcgatgcct cggatgagac
     3901 gaatcgagtt tgaggcaagc tatgcgacac acccggccgc gggtaaccgt ggcggggcat
     3961 ggccgacaaa cagaacgtga aagcgcccaa gatagaaagc cggtagatgc caaccatcca
     4021 gcagctggtc cgcaagggtc gtcgggacaa gatcagtaag gtcaagaccg cggctctgaa
     4081 gggcagcccg cagcgtcgtg gtgtatgcac ccgcgtgtac accaccactc cgaagaagcc
     4141 gaactcggcg cttcggaagg ttgcccgcgt gaagttgacg agtcaggtcg aggtcacggc
     4201 gtacattccc ggcgagggcc acaacctgca ggaggactcg atggtgctgg tgcgcggcgg
     4261 ccgggtgaag gacctgcctg gtgtgcgcta caagatcatc cgcggttcgc tggatacgca
     4321 gggtgtcaag aaccgcaaac aggcacgcag ccgttacggc gctaagaagg agaagggctg
     4381 atgcnacgca agggatctag cccgcctaat gagcgggctt ttttttagat ccccctattt
     4441 aaattcgcga ttaattaaga agttcctata ctttctagag aataggaact tcgagctcga
     4501 attcgataaa ataaaaaagg ggaccctaga ggtccccttt tttattttat cgaattcgtt
     4561 gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt
     4621 aggtggcggt acttgggtcg at
//
