LOCUS       sequence-178581-        4751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSY8 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 4751)
  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 4751)
  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: pSY14-ptb
FEATURES             Location/Qualifiers
     source          1..4751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(356..373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(527..1115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(607..626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1286..2077,2078..2146))
                     /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     1909..1928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2147..2251)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2319..2337
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2375..2397)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2497..2516
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cccttgtcac taagaaaata aatgcagggt aaaatttata tccttcttgt tttatgtttc
       61 ggtataaaac actaatatca atttctgtgg ttatactaaa agtcgtttgt tggttcaaat
      121 aatgattaaa tatctctttt ctcttccaat tgtctaaatc aattttatta aagttcattt
      181 gatatgcctc ctaaattttt atctaaagtg aatttaggag gcttacttgt ctgctttctt
      241 cattagaatc aatccttttt taaaagtcaa tattactgta acataaacag ctgcattaat
      301 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
      361 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      721 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      961 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1201 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1261 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1321 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1381 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1441 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1501 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1561 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1621 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1681 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1741 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1801 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1861 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1921 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1981 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2041 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2101 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2161 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2221 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2281 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     2341 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2401 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2461 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2521 gagtgcacca tatgctcgag ttatttaaga aaagtttcac tactaattgt atttggaagt
     2581 ttatacatca tttgtggaat aagatatggt ttaattaaat tgtaaaatat aagatatgac
     2641 atactatcaa tataaattat aggtttattt ttatttattt ttacataaca attaagttga
     2701 aatttatttc taagtccttt tactaaatat tcaacttctt caaaagtaaa agattgtgta
     2761 tttaaaacta tacttttatt agtagaattt ttattataat cccatttacc tccatcatcc
     2821 ataaaccaat atgcaagtga cattggagta agataatttt ctactaaatt attaggaatt
     2881 gttttcttat tattaacaat aaaaagatta gcaagtttat taaatgcttg atgtttaaaa
     2941 gtttgagcac cccatgtaat tacaagattt ccaagatgat taactctttg ttttttatgt
     3001 ggaggtgaaa gtacccattg atcataaaga agacaaacat gatccatata tgctttattt
     3061 ttccattcaa attgcataca atatgtttta ccttcatctc tacttcttat ataagcatct
     3121 ccaagaataa gaccaattcc tgcttcaaat tgttctatat taagttctat aagttgtgat
     3181 ttatattctt taagaagttt actattagga ccaagattca ttacttgatt cttttgatgc
     3241 atatgtcgac actccctttt actatttaat tatctatgtt aaatgattaa cgtgctcatt
     3301 tatattttaa caaacttttc acatgaagtt aagtttttta gaaaattatt tatattttat
     3361 agaattcccg gggatccgtc gacctgcagc caagcttaat cgcatttcat agattgacct
     3421 cccaataact acgtggtgtt attgggaggt caatctattt catttgcctc ttgctcaaag
     3481 ttcccaaatt cgagtaagag gtatttttgt ttttggtcgt cgcctctcat tagtagttca
     3541 gggtttaaca ttaatactcc agtttttctt tttataatat ttccttcttc taagatttta
     3601 agtgttgtta ttactgtttg tagacttgtt cctgtagctt ttgctatttc tcttgttgta
     3661 gctatcattg tattgttact taagtggaca ttatctagga tatagttaac gattttaagt
     3721 ttttttccgc caatcatatc taacatactt attaattgca ctatatatgc ctttacgaag
     3781 ttaccagacg tttgtttacg gtataacttg tctacctcta tgacttctcc actttcttcg
     3841 tctatgagcc tctgagagcc tttatagact gttccatatc tttctttcat ctttttctca
     3901 ctccttattt taaactattc taactatatc ataactgttc taaaaaaaaa agaacatttg
     3961 ttaaaagaaa ttagaacaaa atgagtgaaa aattagaaca aacaaattcc ttataaacct
     4021 tatcatctca acctatatta agattttacc tagttgaatc ttcttttcta tataaagcgt
     4081 cggagcatat cagggggtta tctaacgtaa atgctaccct tcggctcgct ttcgctcggc
     4141 attgacgtca gatactgcac cccctgaacc cccatgctcc aacagcaaaa aggaaacttt
     4201 ttgctgcttt tccgacgctt attcgcttcg ctcatattta tatagaaaag aagtgaatgc
     4261 gcaaaagaca taatcgattt ataaaagcca gtcattaggc ctatctgaca attcctgaat
     4321 agagttcata aacaatcctg catgataacc atcacaaaca gaatgatgta cctgtaaaga
     4381 tagcggtaaa tatattgaat tacctttatt aatgaatttt cctgctgtaa taatgggtag
     4441 aaggtaatta ctattattat tgatatttaa gttaaaccca gtaaatgaag tccatggaat
     4501 aatagaaaga gaaaaagcat tttcaggtat aggtgttttg ggaaacaatt tccccgaacc
     4561 attatatttc tctacatcag aaaggtataa atcataaaac tctttgaagt cattctttac
     4621 aggagtccaa ataccagaga atgttttaga tacaccatca aaaattgtat aaagtggctc
     4681 taacttatcc caataaccta actctccgtc gctattgtaa ccagttctaa aagctgtatt
     4741 tgagtttatc a
//
