LOCUS       sequence-175410-        6224 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  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 6224)
  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..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..26)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9..25)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(18..40)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(234..253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     353..375
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(413..431)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        499..603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(604..672,673..1464)
                     /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(822..841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1635..2223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2124..2143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2377..2394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2511..2532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2547..2552,2553..2570,2571..2577)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2585..2601
                     /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     2590..2612
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2609..2625
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2609..2625
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gggaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
       61 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      121 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
      181 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
      241 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
      301 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
      361 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
      421 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
      481 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      541 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      601 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      661 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      721 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      781 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      841 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      901 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      961 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1021 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1081 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1141 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1201 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1261 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1321 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1381 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1441 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1501 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1561 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1621 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1681 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1741 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     1801 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1861 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1921 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1981 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2041 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2101 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2161 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2221 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2281 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2341 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2401 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2461 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     2521 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2581 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     2641 ctttggctaa cacacacgcc attccaacca atagttttct cggcataaag ccatgctctg
     2701 acgcttaaat gcactaatgc cttaaaaaaa cattaaagtc taacacacta gacttattta
     2761 cttcgtaatt aagtcgttaa accgtgtgct ctacgaccaa aagtataaaa cctttaagaa
     2821 ctttcttttt tcttgtaaaa aaagaaacta gataaatctc tcatatcttt tattcaataa
     2881 tcgcatcaga ttgcagtata aatttaacga tcactcatca tgttcatatt tatcagagcc
     2941 acaatgaata ggtttacact tactttagtt ttatggaaat gaaagatcat atcatatata
     3001 atctagaata aaattaacta aaataattat tatctagata aaaaatttag aagccaatga
     3061 aatctataaa taaactaaat taagtttatt taattaacaa ctatggatat aaaataggta
     3121 ctaatcaaaa tagtgaggag gatatatttg aatacatacg aacaaattaa taaagtgaaa
     3181 aaaatacttc ggaaacattt aaaaaataac cttattggta cttacatgtt tggatcagga
     3241 gttgagagtg gactaaaacc aaatagtgat cttgactttt tagtcgtcgt atctgaacca
     3301 ttgacagatc aaagtaaaga aatacttata caaaaaatta gacctatttc aaaaaaaata
     3361 ggagataaaa gcaacttacg atatattgaa ttaacaatta ttattcagca agaaatggta
     3421 ccgtggaatc atcctcccaa acaagaattt atttatggag aatggttaca agagctttat
     3481 gaacaaggat acattcctca gaaggaatta aattcagatt taaccataat gctttaccaa
     3541 gcaaaacgaa aaaataaaag aatatacgga aattatgact tagaggaatt actacctgat
     3601 attccatttt ctgatgtgag aagagccatt atggattcgt cagaggaatt aatagataat
     3661 tatcaggatg atgaaaccaa ctctatatta actttatgcc gtatgatttt aactatggac
     3721 acgggtaaaa tcataccaaa agatattgcg ggaaatgcag tggctgaatc ttctccatta
     3781 gaacataggg agagaatttt gttagcagtt cgtagttatc ttggagagaa tattgaatgg
     3841 actaatgaaa atgtaaattt aactataaac tatttaaata acagattaaa aaaattacga
     3901 taatttacag aaaagaaaat tatagaattt agtatgatta attatactca tttatgaatg
     3961 tttaattgaa tacaaaaaaa aatacttgtt atgtattcaa ttacgggtta aaatatagac
     4021 aagttgaaaa atttaataaa aaaataagtc ctcagctctt atatattaag ctaccaactt
     4081 agtatataag ccaaaactta aatgtgctac caacacatca agccgttaga gaactctatc
     4141 tatagcaata tttcaaatgt accgacatac aagagaaaca ttaactatat atattcaatt
     4201 tatgagatta tcttaacaga tataaatgta aattgcaata agtaagattt agaagtttat
     4261 agcctttgtg tattggaagc agtacgcaaa ggctttttta tttgataaaa attagaagta
     4321 tatttatttt ttcataatta atttatgaaa atgaaagggg gtgagcaaag tgacagagga
     4381 aagcagtatc ttatcaaata acaaggtatt agcaatatca ttattgactt tagcagtaaa
     4441 cattatgact tttatagtgc ttgtagctaa gtagtacgaa agggggagct ttaaaaagct
     4501 ccttggaata catagaattc ataaattaat ttatgaaaag aagggcgtat atgaaaactt
     4561 gtaaaaattg caaagagttt attaaagata ctgaaatatg caaaatacat tcgttgatga
     4621 ttcatgataa aacagtagca acctattgca gtaaatacaa tgagtcaaga tgtttacata
     4681 aagggaaagt ccaatgtatt aattgttcaa agatgaaccg atatggatgg tgtgccataa
     4741 aaatgagatg ttttacagag gaagaacaga aaaaagaacg tacatgcatt aaatattatg
     4801 caaggagctt taaaaaagct catgtaaaga agagtaaaaa gaaaaaataa tttatttatt
     4861 aatttaatat tgagagtgcc gacacagtat gcactaaaaa atatatctgt ggtgtagtga
     4921 gccgatacaa aaggatagtc actcgcattt tcataataca tcttatgtta tgattatgtg
     4981 tcggtgggac ttcacgacga aaacccacaa taaaaaaaga gttcggggta gggttaagca
     5041 tagttgaggc aactaaacaa tcaagctagg atatgcagta gcagaccgta aggtcgttgt
     5101 ttaggtgtgt tgtaatacat acgctattaa gatgtaaaaa tacggatacc aatgaaggga
     5161 aaagtataat ttttggatgt agtttgtttg ttcatctatg ggcaaactac gtccaaagcc
     5221 gtttccaaat ctgctaaaaa gtatatcctt tctaaaatca aagtcaagta tgaaatcata
     5281 aataaagttt aattttgaag ttattatgat attatgtttt tctattaaaa taaattaagt
     5341 atatagaata gtttaataat agtatatact taatgtgctg cagtataaaa tataaataat
     5401 tttctaaaaa acttaacttc atgtgaaaag tttgttaaaa tataaatgag cacgttaatc
     5461 atttaacata gataattaaa tagtaaaagg gagtgtcgac atatgcatca aaagaatcaa
     5521 gtaatgaatc ttggtcctaa tagtaaactt cttaaagaat ataaatcaca acttatagaa
     5581 cttaatatag aacaatttga agcaggaatt ggtcttattc ttggagatgc ttatataaga
     5641 agtagagatg aaggtaaaac atattgtatg caatttgaat ggaaaaataa agcatatatg
     5701 gatcatgttt gtcttcttta tgatcaatgg gtactttcac ctccacataa aaaacaaaga
     5761 gttaatcatc ttggaaatct tgtaattaca tggggtgctc aaacttttaa acatcaagca
     5821 tttaataaac ttgctaatct ttttattgtt aataataaga aaacaattcc taataattta
     5881 gtagaaaatt atcttactcc aatgtcactt gcatattggt ttatggatga tggaggtaaa
     5941 tgggattata ataaaaattc tactaataaa agtatagttt taaatacaca atcttttact
     6001 tttgaagaag ttgaatattt agtaaaagga cttagaaata aatttcaact taattgttat
     6061 gtaaaaataa ataaaaataa acctataatt tatattgata gtatgtcata tcttatattt
     6121 tacaatttaa ttaaaccata tcttattcca caaatgatgt ataaacttcc aaatacaatt
     6181 agtagtgaaa cttttcttaa ataataaaag taattacatt accc
//
