LOCUS       sequence-36462-e        7987 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 7987)
  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 7987)
  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: pMN234
FEATURES             Location/Qualifiers
     source          1..7987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(28..1299)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     terminator      complement(1483..1510)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      1639..2227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2128..2147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2381..2398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2413..2553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2479..2498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2639..2661
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2655..2846)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             3693..4487
                     /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(3747..3766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4357..4376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7924..7940
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7925..7941
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcttgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta
       61 gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc
      121 actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa
      181 tgctatcatt tcctttgata ttggatcata tgcatagtac cgagaaacta gtgcgaagta
      241 gtgatcaggt attgctgtta tctgatgagt atacgttgtc ctggccacgg cagaagcacg
      301 cttatcgctc caatttccca caacattagt caactccgtt aggcccttca ttgacagaaa
      361 tgaggtcatc aaatgtcttc caatgtgaga ttttgggcca ttctttatag caaagattgg
      421 ataaggcgca tttttcttca aagccttgtt gtacgatctg actaagttat cttttaataa
      481 ttggtattcc tgtttgttgc ttgaagaatt gccggtccta tttactcgtt ttaggactgg
      541 ttcagaattc ctcaaaaatt catccaaata tacaagtgga tcgatcctac cccttgcgct
      601 aaagaagtat atgtgcctac taacgcttgt ctttgtctct gtcactaaac actggattat
      661 tactcccaga tacttatttt ggactaattt aaatgatttc ggatcaacgt tcttaatatc
      721 gctgaatctt ccacaattga tgaaagtagc taggaagagg aattggtata aagtttttgt
      781 ttttgtaaat ctcgaggtat actcaaacga atttagtatt ttctcagtga tctcccagat
      841 gctttcaccc tcacttagaa gtgctttaag cattttttta ctgtggctat ttcccttatc
      901 tgcttcttcc gatgattcga actgtaattg caaactactt acaatatcag tgatatcaga
      961 ttgatgtttt tgtccattgt aaggaataat tgtaaattcc caagcaggaa ttaatttctt
     1021 taatgaggct tccagaattg ttgctttttg cgtcttgtat ttaaactgga gtgatttgtt
     1081 gacaatatcg aaactcagcg aattgcttat gatagtatta tagctcatga atgtggctct
     1141 cttgattgct gttccgttat gagtaatcat ccaacataaa taggttagtt cagcagcaca
     1201 tgatgctatt ttttcccctg aaggtctttc aaacctttcc acaaactgac gaaccaggac
     1261 cttaggtggt gttttacata atatatcaaa ttgtggcata ttaacctcct ttctgttaat
     1321 taagcatgcg gatccgtggc angagggtcg cctgcgtcgg caactcgccc aggctacacg
     1381 agagccctcg actgcgaaaa tgccgaccag gcaccaccgc actccaggct taacccggtg
     1441 cgacgatcca atatttctag agtttaaaca gtattaaacg caaaaaggcc atccgtcagg
     1501 atggccttct gcttagctaa tcaactagaa aaggatctag gtgaagatcc tttttgataa
     1561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1741 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2341 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2401 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2461 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     2521 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     2581 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2641 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     2701 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     2761 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     2821 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     2881 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     2941 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3001 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     3061 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     3121 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     3181 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     3241 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     3301 atgcgcaccc gtggccagga cccaacgctg cccgacgact tcacgctgcc gcaagcactc
     3361 agggcgcaag ggctgctaaa aggaagcgga acacgtagaa agccagtccg cagaaacggt
     3421 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     3481 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     3541 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     3601 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat
     3661 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3721 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3781 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3841 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3901 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3961 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4021 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4081 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4141 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4201 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4261 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4321 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4381 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4441 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4501 taccgcgtca acgtagcgcc ctgcttcggc cgcaacgccc gctttgacct gccagactgg
     4561 cggcgggtat tgtggttgct cgtgcctggc ggcttacgct tgatgtaggg gcgtttgatg
     4621 ccgggccaat tcgcatgtcc gcgatgcctc ggatgagacg aatcgagttt gaggcaagct
     4681 atgcgacaca cccggccgcg ggtaaccgtg gcggggcatg gccgacaaac agaacgtgaa
     4741 agcgcccaag atagaaagcc ggtagatgcc aaccatccag cagctggtcc gcaagggtcg
     4801 tcgggacaag atcagtaagg tcaagaccgc ggctctgaag ggcagcccgc agcgtcgtgg
     4861 tgtatgcacc cgcgtgtaca ccaccactcc gaagaagccg aactcggcgc ttcggaaggt
     4921 tgcccgcgtg aagttgacga gtcaggtcga ggtcacggcg tacattcccg gcgagggcca
     4981 caacctgcag gaggactcga tggtgctggt gcgcggcggc cgggtgaagg acctgcctgg
     5041 tgtgcgctac aagatcatcc gcggttcgct ggatacgcag ggtgtcaaga accgcaaaca
     5101 ggcacgcagc cgttacggcg ctaagaagga gaagggctga tgcnacgcaa gggatctagc
     5161 ccgcctaatg agcgggcttt tttttagatc ccccccgacg gcggtgcggg cccggcggcg
     5221 gacgctcagc agcggcgggc gtgaaaggcc ctggcatcct cgatcatctc ctccagggtg
     5281 gtcggccgag cttccatccc agctcggcaa ggatcaacga ggacagtcgc acgacgaagt
     5341 tcttctggat cgcgcccgtg ctggaagcac tcaacctcga agcgtgtggt tgcggagcca
     5401 tctagcaacc acacgaaaca tgcgcaacga accgcgcaac gaacaacgcc tagaactggc
     5461 cctagatgag ctgactcgta tcgttggtaa acctagtttg accagcatgt tttaactacg
     5521 ttcggtgagc tgtcaacggg gcctgtaacg gcacaacgaa ccgtgcaacg agagtggcca
     5581 cggatgccac cacaggcact acaacggagt tcgccacgta catcaccaca accaccgatt
     5641 ctggcggtga gctccccgat attcagcgga aatggcttgg tatcgaccaa gattcgtaga
     5701 accccgtctc gtctggctgg tattcaaaac gggcgcaacg aaacacgcaa cgagacaggc
     5761 atggcccaaa ccagaaaact agcgtctacc aggactttta cctgtccgac ccgttgcaac
     5821 ggaacccccc acggaacccc cgcgacaccc gctccccaat tgcgttagaa cagcggtgga
     5881 ttgtcggctt cgttgtgggc cttttgagcc gcttcctgtt ctgccgcacg ctctttcctc
     5941 gcccgatagc cgagtcgctt aacggtgtcc agatgcagcc cgaaatgttt ggccgtttgc
     6001 ggccaagagt ggccctcgtc gtcgtgatag gcgcggatgc gttcgcggcg tgcagcctgc
     6061 tcggcgagcc actcgctgcg ttcctgcgcc acgagccgga cgacgtggcg ttcggatagt
     6121 ccggtgattc gagcgccttc ggcggcggtc acgcgccgct ttttgcggac agtcggctgc
     6181 cggttgtagc cgtcgctgta gccgtcgctg tagccgtcgc tcatagcaat gcctccatgg
     6241 ctgacgcgga ctttgcgcgc cgcgcaactg tgctcgccgc cgtgcgcgct gctgcgccct
     6301 tccgcgagat ggccgactgg cgcgcactga gtgtggcctc gtagaccacg atcccgtccg
     6361 cccaaatgcg cgacttggtt gtgatccaac gccaaatgct gttggcgatg gcgcggacct
     6421 cgctgtccgg tagcggtccg ggacacacgt cgttgcacgg gaattcggcg tttcgcgcgt
     6481 ggcactcggc atagatcgcg cggccgagtc cgtccacgtt ccgggtcggc aggtagatcc
     6541 gcatgagggc gggacgatag gcccacaacc tgacggaatc gaacagtgcg caattccgcc
     6601 ctagcggcgt cggagccgct ttgtacgtgg tctgctgacg ccagcgcggc ggtggcatgt
     6661 tcgcgccgag ctcggcctcg atgtggctga gtgtgtagag atctgagtgg agccattccg
     6721 tttcccaggc gatgtggccg gggtttttgg tcatgaggcc tgagtaactg cggtcgccgt
     6781 cgacggcgcg ccgaaggcct tcggcgcacg ccgccatgta tgcgagcggc ttacgccgcg
     6841 cgtattcggt gcgtggaaca ggggcgttga gtgcccacac tgcgtgtgcg tggccgttgg
     6901 cgcgattgcc cacgatcgcg ttgggcagcg gatgggaccc ccgggcgctg agcgctcgga
     6961 gcgctgcgtc tggatggtct acgtccacga ccagcaggtt tgccagcgct gttgggttcg
     7021 cctcgatgta ccggcggcct agggccgacg cgcggctttg gcggtagatc ccctcgagca
     7081 gatcgtcgct tgccagcggc cagtacggca gccagagctg ctcaaattcg tcggcgacgt
     7141 ggctcacgct tggtagtaga ccacgattaa tcaccggtgt atggtccgac acgagctcca
     7201 agtcagatat ttcgctgagg ggccacccca caactgcaca ctcccccgct ctcccgtcga
     7261 gccctggtgg tggaacacca gcgacagccg agcaccccca accacctgta ccaaccagga
     7321 ggaacacatg cgtcgtttcg aggacgtttc cgggccgctg agagccgctg tggcggccgt
     7381 acacgccgcc ttagacccgt tagaccccct gccgcctgaa tgcgcgggta cgagccacac
     7441 agcgcccgaa cttacggagc tggtgggctc acctggcttt atggcgtacg aatcggctgt
     7501 gtgcgacctg ttgggcgagg tgaggtacgc gctactcacg ctggcaaggg cgacacagcc
     7561 gccccaccga gcccgcacgg ccgcgcgcgg tgtcaacaac cgggtgagtc gtgcacacca
     7621 gcaggtgttc gaggcttggc tcgaagtgca ggacatcgtg gcgaacgccg cccgatgagc
     7681 cgcgccttac gctggctgcc agccgttcgc gggctggttg gtgcagcgcg tcgagcggtt
     7741 agaggccctg cggtgttcca ccaccgcagg cctcgccctt tttaaggctg aatttgcttg
     7801 tctccgaatc caactggctt gtccaagggt gtatctacgc ttaatccaaa gttcaaacga
     7861 ggggattaca catgaccaac ttcgataacg ttctcggctc gatgatcccc ggcgggcccc
     7921 ccctcgaggt cgacggtatc gataaaataa aaaaggggac ctctagggtc cccaattaat
     7981 tagctaa
//
