LOCUS       sequence-36551-e        7968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses M. tuberculosis codon-optimized flpe gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7968)
  AUTHORS   Song H, Niederweis M
  TITLE     Functional expression of the Flp recombinase in Mycobacterium bovis 
            BCG.
  JOURNAL   Gene. 2007 Sep 15;399(2):112-9. Epub 2007 May 21.
  PUBMED    17590537
REFERENCE   2  (bases 1 to 7968)
  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: "2007-09-15"; volume: "399(2)"; pages: "112-9. Epub 2007 May 
            21"
COMMENT     Sequence Label: pML597
FEATURES             Location/Qualifiers
     source          1..7968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(153..180)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      309..897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     798..817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1051..1068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1083..1223
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1149..1168)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1309..1331
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1325..1516)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             2363..3157
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2417..2436)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3027..3046
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6594..6610
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6595..6611
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gatccgtggc angagggtcg cctgcgtcgg caactcgccc aggctacacg agagccctcg
       61 actgcgaaaa tgccgaccag gcaccaccgc actccaggct taacccggtg cgacgatcca
      121 atatttctag agtttaaaca gtattaaacg caaaaaggcc atccgtcagg atggccttct
      181 gcttagctaa tcaactagaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      421 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
      481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1021 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1081 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1141 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     1201 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     1261 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     1321 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     1381 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     1441 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     1501 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     1561 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     1621 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     1681 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     1741 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     1801 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     1861 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     1921 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     1981 gtggccagga cccaacgctg cccgacgact tcacgctgcc gcaagcactc agggcgcaag
     2041 ggctgctaaa aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg
     2101 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
     2161 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
     2221 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     2281 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
     2341 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2401 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2461 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2521 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     2581 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2641 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2701 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2761 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     2821 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2881 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2941 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3001 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3061 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3121 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg taccgcgtca
     3181 acgtagcgcc ctgcttcggc cgcaacgccc gctttgacct gccagactgg cggcgggtat
     3241 tgtggttgct cgtgcctggc ggcttacgct tgatgtaggg gcgtttgatg ccgggccaat
     3301 tcgcatgtcc gcgatgcctc ggatgagacg aatcgagttt gaggcaagct atgcgacaca
     3361 cccggccgcg ggtaaccgtg gcggggcatg gccgacaaac agaacgtgaa agcgcccaag
     3421 atagaaagcc ggtagatgcc aaccatccag cagctggtcc gcaagggtcg tcgggacaag
     3481 atcagtaagg tcaagaccgc ggctctgaag ggcagcccgc agcgtcgtgg tgtatgcacc
     3541 cgcgtgtaca ccaccactcc gaagaagccg aactcggcgc ttcggaaggt tgcccgcgtg
     3601 aagttgacga gtcaggtcga ggtcacggcg tacattcccg gcgagggcca caacctgcag
     3661 gaggactcga tggtgctggt gcgcggcggc cgggtgaagg acctgcctgg tgtgcgctac
     3721 aagatcatcc gcggttcgct ggatacgcag ggtgtcaaga accgcaaaca ggcacgcagc
     3781 cgttacggcg ctaagaagga gaagggctga tgcnacgcaa gggatctagc ccgcctaatg
     3841 agcgggcttt tttttagatc ccccccgacg gcggtgcggg cccggcggcg gacgctcagc
     3901 agcggcgggc gtgaaaggcc ctggcatcct cgatcatctc ctccagggtg gtcggccgag
     3961 cttccatccc agctcggcaa ggatcaacga ggacagtcgc acgacgaagt tcttctggat
     4021 cgcgcccgtg ctggaagcac tcaacctcga agcgtgtggt tgcggagcca tctagcaacc
     4081 acacgaaaca tgcgcaacga accgcgcaac gaacaacgcc tagaactggc cctagatgag
     4141 ctgactcgta tcgttggtaa acctagtttg accagcatgt tttaactacg ttcggtgagc
     4201 tgtcaacggg gcctgtaacg gcacaacgaa ccgtgcaacg agagtggcca cggatgccac
     4261 cacaggcact acaacggagt tcgccacgta catcaccaca accaccgatt ctggcggtga
     4321 gctccccgat attcagcgga aatggcttgg tatcgaccaa gattcgtaga accccgtctc
     4381 gtctggctgg tattcaaaac gggcgcaacg aaacacgcaa cgagacaggc atggcccaaa
     4441 ccagaaaact agcgtctacc aggactttta cctgtccgac ccgttgcaac ggaacccccc
     4501 acggaacccc cgcgacaccc gctccccaat tgcgttagaa cagcggtgga ttgtcggctt
     4561 cgttgtgggc cttttgagcc gcttcctgtt ctgccgcacg ctctttcctc gcccgatagc
     4621 cgagtcgctt aacggtgtcc agatgcagcc cgaaatgttt ggccgtttgc ggccaagagt
     4681 ggccctcgtc gtcgtgatag gcgcggatgc gttcgcggcg tgcagcctgc tcggcgagcc
     4741 actcgctgcg ttcctgcgcc acgagccgga cgacgtggcg ttcggatagt ccggtgattc
     4801 gagcgccttc ggcggcggtc acgcgccgct ttttgcggac agtcggctgc cggttgtagc
     4861 cgtcgctgta gccgtcgctg tagccgtcgc tcatagcaat gcctccatgg ctgacgcgga
     4921 ctttgcgcgc cgcgcaactg tgctcgccgc cgtgcgcgct gctgcgccct tccgcgagat
     4981 ggccgactgg cgcgcactga gtgtggcctc gtagaccacg atcccgtccg cccaaatgcg
     5041 cgacttggtt gtgatccaac gccaaatgct gttggcgatg gcgcggacct cgctgtccgg
     5101 tagcggtccg ggacacacgt cgttgcacgg gaattcggcg tttcgcgcgt ggcactcggc
     5161 atagatcgcg cggccgagtc cgtccacgtt ccgggtcggc aggtagatcc gcatgagggc
     5221 gggacgatag gcccacaacc tgacggaatc gaacagtgcg caattccgcc ctagcggcgt
     5281 cggagccgct ttgtacgtgg tctgctgacg ccagcgcggc ggtggcatgt tcgcgccgag
     5341 ctcggcctcg atgtggctga gtgtgtagag atctgagtgg agccattccg tttcccaggc
     5401 gatgtggccg gggtttttgg tcatgaggcc tgagtaactg cggtcgccgt cgacggcgcg
     5461 ccgaaggcct tcggcgcacg ccgccatgta tgcgagcggc ttacgccgcg cgtattcggt
     5521 gcgtggaaca ggggcgttga gtgcccacac tgcgtgtgcg tggccgttgg cgcgattgcc
     5581 cacgatcgcg ttgggcagcg gatgggaccc ccgggcgctg agcgctcgga gcgctgcgtc
     5641 tggatggtct acgtccacga ccagcaggtt tgccagcgct gttgggttcg cctcgatgta
     5701 ccggcggcct agggccgacg cgcggctttg gcggtagatc ccctcgagca gatcgtcgct
     5761 tgccagcggc cagtacggca gccagagctg ctcaaattcg tcggcgacgt ggctcacgct
     5821 tggtagtaga ccacgattaa tcaccggtgt atggtccgac acgagctcca agtcagatat
     5881 ttcgctgagg ggccacccca caactgcaca ctcccccgct ctcccgtcga gccctggtgg
     5941 tggaacacca gcgacagccg agcaccccca accacctgta ccaaccagga ggaacacatg
     6001 cgtcgtttcg aggacgtttc cgggccgctg agagccgctg tggcggccgt acacgccgcc
     6061 ttagacccgt tagaccccct gccgcctgaa tgcgcgggta cgagccacac agcgcccgaa
     6121 cttacggagc tggtgggctc acctggcttt atggcgtacg aatcggctgt gtgcgacctg
     6181 ttgggcgagg tgaggtacgc gctactcacg ctggcaaggg cgacacagcc gccccaccga
     6241 gcccgcacgg ccgcgcgcgg tgtcaacaac cgggtgagtc gtgcacacca gcaggtgttc
     6301 gaggcttggc tcgaagtgca ggacatcgtg gcgaacgccg cccgatgagc cgcgccttac
     6361 gctggctgcc agccgttcgc gggctggttg gtgcagcgcg tcgagcggtt agaggccctg
     6421 cggtgttcca ccaccgcagg cctcgccctt tttaaggctg aatttgcttg tctccgaatc
     6481 caactggctt gtccaagggt gtatctacgc ttaatccaaa gttcaaacga ggggattaca
     6541 catgaccaac ttcgataacg ttctcggctc gatgatcccc ggcgggcccc ccctcgaggt
     6601 cgacggtatc gataaaataa aaaaggggac ctctagggtc cccaattaat tagctaaagc
     6661 ttatttaaat tcagatgcgg cggttgatgt acgacgacag gtagtccagc acctcctgcg
     6721 agatgatgcc gttccaggcc gggtagcgga tcgagccctc ggccgagccc ttcagctgct
     6781 cgatgtgctg ccactcctcg atcgggttgg tctcgtcctt cagggcgatc atctccttcg
     6841 agatcgggtc gtaggcgtag tagcgcgaca ccagggcgaa gtagtggtcc gggatggcgg
     6901 tgatctggtg ggtgtaggtc gtccgggcca cggccgaggc gcgcttgtcc gaccagttgc
     6961 ccaccacgtt ggtcagctcg gtcaggccct tcatcgacag gaacgaggtc atcaggtggc
     7021 ggccgatgtg cgacttcggg ccgttcttga tggcgaagat cgggtacggg gcgttcttct
     7081 tcagggcctt gttgtacgag cgcaccaggt tgtccttcag cagctggtac tcctgcttgt
     7141 tcgacgacga gttgccggtg cggttcacgc gcttcagcac cggctccgag ttccgcagga
     7201 actcgtccag gtacaccagc gggtcgatgc ggccgcgcgc cgagaagaag tagatgtggc
     7261 gcgacaccga ggtcttggtc tcggtcacga ggcactggat gatcacgccc aggtacttgt
     7321 tctgcaccag cttgaacgac ttcgggtcca cgttcttgat gtccgagaag cggccgcagt
     7381 tgatgaaggt ggccaggaac aggaactggt acagggtctt ggtcttggtg aagcgcgagg
     7441 tgtactcgaa cgagttcagg atcttctcgg tgatctccca gatcgactcg ccctccgaca
     7501 gcagggcctt cagcatcttc ttcgagtgcg agttgccctt gtcggcctcc tccgacgact
     7561 cgaactgcag ctgcagcgac gacacgatgt cggtgatgtc cgactggtgc ttctggccgt
     7621 tgtacgggat gatggtgaac tcccaggccg ggatcagctt cttcagcgac gcctccagga
     7681 tggtggcctt ctgggtcttg tacttgaact gcagcgactt gttcacgatg tcgaacgaca
     7741 gcgagttcga gatgatggtg ttgtacgaca tgaaggtggc gcgcttgatg gccgtgccgt
     7801 tgtgggtgat catccagcac aggtaggtca gctcggcggc gcacgaggcg atcttctcgc
     7861 ccgacgggcg ctcgaagcgc tccacgaact ggcgcaccag caccttcggc ggggtcttgc
     7921 acaggatgtc gaactgcggc attattaacc tcctttctgt taattaag
//
