LOCUS       sequence-181274-        6109 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pXY1-spc derivative with sceC gene under the control of ptb promoter
            used for I-SceI-mediated scarless gene modification via allelic 
            exchange in Clostridium.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6109)
  AUTHORS   Zhang N, Shao L, Jiang Y, Gu Y, Li Q, Liu J, Jiang W, Yang S
  TITLE     I-SceI-mediated scarless gene modification via allelic exchange in 
            Clostridium.
  JOURNAL   J Microbiol Methods. 2015 Jan;108:49-60. doi: 
            10.1016/j.mimet.2014.11.004. Epub 2014 Nov 15.
  PUBMED    25451462
REFERENCE   2  (bases 1 to 6109)
  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: "J Microbiol
            Methods."; date: "2015-01"; volume: "108:49-60. doi"; pages: " 
            10.1016/j.mimet.2014.11.004. Epub 2014 Nov 15"
COMMENT     Sequence Label: pXY1-spc-ptb
FEATURES             Location/Qualifiers
     source          1..6109
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(212..231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     331..353
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(391..409)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        477..581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(582..650,651..1442)
                     /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     complement(800..819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1613..2201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2102..2121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2355..2372
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2489..2510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2525..2530,2531..2548,2549..2555)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2563..2579
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2568..2590
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2587..2603
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2587..2603
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
       61 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      121 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
      181 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
      241 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
      301 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
      361 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
      421 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      481 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      541 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      601 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      661 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      721 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      781 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      841 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      901 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      961 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1021 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1081 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1141 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1201 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1261 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1321 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1381 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1441 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1501 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1561 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1621 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1681 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1741 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1801 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1861 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1921 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1981 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2041 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2101 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2161 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2221 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2281 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2341 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     2401 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2461 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     2521 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     2581 ttcacacagg aaacagctat gaccatgatt acgccaagct ttggctaaca cacacgccat
     2641 tccaaccaat agttttctcg gcataaagcc atgctctgac gcttaaatgc actaatgcct
     2701 taaaaaaaca ttaaagtcta acacactaga cttatttact tcgtaattaa gtcgttaaac
     2761 cgtgtgctct acgaccaaaa gtataaaacc tttaagaact ttcttttttc ttgtaaaaaa
     2821 agaaactaga taaatctctc atatctttta ttcaataatc gcatcagatt gcagtataaa
     2881 tttaacgatc actcatcatg ttcatattta tcagagccac aatgaatagg tttacactta
     2941 ctttagtttt atggaaatga aagatcatat catatataat ctagaataaa attaactaaa
     3001 ataattatta tctagataaa aaatttagaa gccaatgaaa tctataaata aactaaatta
     3061 agtttattta attaacaact atggatataa aataggtact aatcaaaata gtgaggagga
     3121 tatatttgaa tacatacgaa caaattaata aagtgaaaaa aatacttcgg aaacatttaa
     3181 aaaataacct tattggtact tacatgtttg gatcaggagt tgagagtgga ctaaaaccaa
     3241 atagtgatct tgacttttta gtcgtcgtat ctgaaccatt gacagatcaa agtaaagaaa
     3301 tacttataca aaaaattaga cctatttcaa aaaaaatagg agataaaagc aacttacgat
     3361 atattgaatt aacaattatt attcagcaag aaatggtacc gtggaatcat cctcccaaac
     3421 aagaatttat ttatggagaa tggttacaag agctttatga acaaggatac attcctcaga
     3481 aggaattaaa ttcagattta accataatgc tttaccaagc aaaacgaaaa aataaaagaa
     3541 tatacggaaa ttatgactta gaggaattac tacctgatat tccattttct gatgtgagaa
     3601 gagccattat ggattcgtca gaggaattaa tagataatta tcaggatgat gaaaccaact
     3661 ctatattaac tttatgccgt atgattttaa ctatggacac gggtaaaatc ataccaaaag
     3721 atattgcggg aaatgcagtg gctgaatctt ctccattaga acatagggag agaattttgt
     3781 tagcagttcg tagttatctt ggagagaata ttgaatggac taatgaaaat gtaaatttaa
     3841 ctataaacta tttaaataac agattaaaaa aattaatcga taatttacag aaaagaaaat
     3901 tatagaattt agtatgatta attatactca tttatgaatg tttaattgaa tacaaaaaaa
     3961 aatacttgtt atgtattcaa ttacgggtta aaatatagac aagttgaaaa atttaataaa
     4021 aaaataagtc ctcagctctt atatattaag ctaccaactt agtatataag ccaaaactta
     4081 aatgtgctac caacacatca agccgttaga gaactctatc tatagcaata tttcaaatgt
     4141 accgacatac aagagaaaca ttaactatat atattcaatt tatgagatta tcttaacaga
     4201 tataaatgta aattgcaata agtaagattt agaagtttat agcctttgtg tattggaagc
     4261 agtacgcaaa ggctttttta tttgataaaa attagaagta tatttatttt ttcataatta
     4321 atttatgaaa atgaaagggg gtgagcaaag tgacagagga aagcagtatc ttatcaaata
     4381 acaaggtatt agcaatatca ttattgactt tagcagtaaa cattatgact tttatagtgc
     4441 ttgtagctaa gtagtacgaa agggggagct ttaaaaagct ccttggaata catagaattc
     4501 ataaattaat ttatgaaaag aagggcgtat atgaaaactt gtaaaaattg caaagagttt
     4561 attaaagata ctgaaatatg caaaatacat tcgttgatga ttcatgataa aacagtagca
     4621 acctattgca gtaaatacaa tgagtcaaga tgtttacata aagggaaagt ccaatgtatt
     4681 aattgttcaa agatgaaccg atatggatgg tgtgccataa aaatgagatg ttttacagag
     4741 gaagaacaga aaaaagaacg tacatgcatt aaatattatg caaggagctt taaaaaagct
     4801 catgtaaaga agagtaaaaa gaaaaaataa tttatttatt aatttaatat tgagagtgcc
     4861 gacacagtat gcactaaaaa atatatctgt ggtgtagtga gccgatacaa aaggatagtc
     4921 actcgcattt tcataataca tcttatgtta tgattatgtg tcggtgggac ttcacgacga
     4981 aaacccacaa taaaaaaaga gttcggggta gggttaagca tagttgaggc aactaaacaa
     5041 tcaagctagg atatgcagta gcagaccgta aggtcgttgt ttaggtgtgt tgtaatacat
     5101 acgctattaa gatgtaaaaa tacggatacc aatgaaggga aaagtataat ttttggatgt
     5161 agtttgtttg ttcatctatg ggcaaactac gtccaaagcc gtttccaaat ctgctaaaaa
     5221 gtatatcctt tctaaaatca aagtcaagta tgaaatcata aataaagttt aattttgaag
     5281 ttattatgat attatgtttt tctattaaaa taaattaagt atatagaata gtttaataat
     5341 agtatatact taawrwrcys cmgtataaaa tataaataat tttctaaaaa acttaacttc
     5401 atgtgaaaag tttgttaaaa tataaatgag cacgttaatc atttaacata gataattaaa
     5461 tagtaaaagg gagtgtcgac atatgcatca aaagaatcaa gtaatgaatc ttggtcctaa
     5521 tagtaaactt cttaaagaat ataaatcaca acttatagaa cttaatatag aacaatttga
     5581 agcaggaatt ggtcttattc ttggagatgc ttatataaga agtagagatg aaggtaaaac
     5641 atattgtatg caatttgaat ggaaaaataa agcatatatg gatcatgttt gtcttcttta
     5701 tgatcaatgg gtactttcac ctccacataa aaaacaaaga gttaatcatc ttggaaatct
     5761 tgtaattaca tggggtgctc aaacttttaa acatcaagca tttaataaac ttgctaatct
     5821 ttttattgtt aataataaga aaacaattcc taataattta gtagaaaatt atcttactcc
     5881 aatgtcactt gcatattggt ttatggatga tggaggtaaa tgggattata ataaaaattc
     5941 tactaataaa agtatagttt taaatacaca atcttttact tttgaagaag ttgaatattt
     6001 agtaaaagga cttagaaata aatttcaact taattgttat gtaaaaataa ataaaaataa
     6061 acctataatt tatattgata gtatgtcata tcttatattt tacaattta
//
