LOCUS       sequence-178610-        5286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with ptb promoter for gene expression in Clostridium 
            acetobutylicum ATCC 824 and Clostridium beijerinckii NCIMB8052.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5286)
  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 5286)
  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
FEATURES             Location/Qualifiers
     source          1..5286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(126..714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(206..225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(885..1676,1677..1745))
                     /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     1508..1527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1746..1850)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1918..1936
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1974..1996)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2096..2115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5010..5026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5010..5026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5023..5045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5034..5050
                     /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)."
     promoter        complement(join(5058..5064,5065..5082,5083..5088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5103..5124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5241..5258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
       61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1861 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     1921 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     1981 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2041 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     2101 tcagagcaga ttgtactgag agtgcaccat atgtcgacac tcccttttac tatttaatta
     2161 tctatgttaa atgattaacg tgctcattta tattttaaca aacttttcac atgaagttaa
     2221 gttttttaga aaattattta tattttatac tgcagcacat taagtatata ctattattaa
     2281 actattctat atacttaatt tattttaata gaaaaacata atatcataat aacttcaaaa
     2341 ttaaacttta tttatgattt catacttgac tttgatttta gaaaggatat actttttagc
     2401 agatttggaa acggctttgg acgtagtttg cccatagatg aacaaacaaa ctacatccaa
     2461 aaattatact tttcccttca ttggtatccg tatttttaca tcttaatagc gtatgtatta
     2521 caacacacct aaacaacgac cttacggtct gctactgcat atcctagctt gattgtttag
     2581 ttgcctcaac tatgcttaac cctaccccga actctttttt tattgtgggt tttcgtcgtg
     2641 aagtcccacc gacacataat cataacataa gatgtattat gaaaatgcga gtgactatcc
     2701 ttttgtatcg gctcactaca ccacagatat attttttagt gcatactgtg tcggcactct
     2761 caatattaaa ttaataaata aattattttt tctttttact cttctttaca tgagcttttt
     2821 taaagctcct tgcataatat ttaatgcatg tacgttcttt tttctgttct tcctctgtaa
     2881 aacatctcat ttttatggca caccatccat atcggttcat ctttgaacaa ttaatacatt
     2941 ggactttccc tttatgtaaa catcttgact cattgtattt actgcaatag gttgctactg
     3001 ttttatcatg aatcatcaac gaatgtattt tgcatatttc agtatcttta ataaactctt
     3061 tgcaattttt acaagttttc atatacgccc ttcttttcat aaattaattt atgaattcta
     3121 tgtattccaa ggagcttttt aaagctcccc ctttcgtact acttagctac aagcactata
     3181 aaagtcataa tgtttactgc taaagtcaat aatgatattg ctaatacctt gttatttgat
     3241 aagatactgc tttcctctgt cactttgctc accccctttc attttcataa attaattatg
     3301 aaaaaataaa tatacttcta atttttatca aataaaaaag cctttgcgta ctgcttccaa
     3361 tacacaaagg ctataaactt ctaaatctta cttattgcaa tttacattta tatctgttaa
     3421 gataatctca taaattgaat atatatagtt aatgtttctc ttgtatgtcg gtacatttga
     3481 aatattgcta tagatagagt tctctaacgg cttgatgtgt tggtagcaca tttaagtttt
     3541 ggcttatata ctaagttggt agcttaatat ataagagctg aggacttatt tttttattaa
     3601 atttttcaac ttgtctatat tttaacccgt aattgaatac ataacaagta tttttttttg
     3661 tattcaatta aacattcata aatgagtata attaatcata ctaaattcta taattttctt
     3721 ttctgtaaat tatcgattaa tttttttaat ctgttattta aatagtttat agttaaattt
     3781 acattttcat tagtccattc aatattctct ccaagataac tacgaactgc taacaaaatt
     3841 ctctccctat gttctaatgg agaagattca gccactgcat ttcccgcaat atcttttggt
     3901 atgattttac ccgtgtccat agttaaaatc atacggcata aagttaatat agagttggtt
     3961 tcatcatcct gataattatc tattaattcc tctgacgaat ccataatggc tcttctcaca
     4021 tcagaaaatg gaatatcagg tagtaattcc tctaagtcat aatttccgta tattctttta
     4081 ttttttcgtt ttgcttggta aagcattatg gttaaatctg aatttaattc cttctgagga
     4141 atgtatcctt gttcataaag ctcttgtaac cattctccat aaataaattc ttgtttggga
     4201 ggatgattcc acggtaccat ttcttgctga ataataattg ttaattcaat atatcgtaag
     4261 ttgcttttat ctcctatttt ttttgaaata ggtctaattt tttgtataag tatttcttta
     4321 ctttgatctg tcaatggttc agatacgacg actaaaaagt caagatcact atttggtttt
     4381 agtccactct caactcctga tccaaacatg taagtaccaa taaggttatt ttttaaatgt
     4441 ttccgaagta tttttttcac tttattaatt tgttcgtatg tattcaaata tatcctcctc
     4501 actattttga ttagtaccta ttttatatcc atagttgtta attaaataaa cttaatttag
     4561 tttatttata gatttcattg gcttctaaat tttttatcta gataataatt attttagtta
     4621 attttattct agattatata tgatatgatc tttcatttcc ataaaactaa agtaagtgta
     4681 aacctattca ttgtggctct gataaatatg aacatgatga gtgatcgtta aatttatact
     4741 gcaatctgat gcgattattg aataaaagat atgagagatt tatctagttt ctttttttac
     4801 aagaaaaaag aaagttctta aaggttttat acttttggtc gtagagcaca cggtttaacg
     4861 acttaattac gaagtaaata agtctagtgt gttagacttt aatgtttttt taaggcatta
     4921 gtgcatttaa gcgtcagagc atggctttat gccgagaaaa ctattggttg gaatggcgtg
     4981 tgtgttagcc aaagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5041 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5101 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5161 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5221 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5281 cgagcg
//
