LOCUS       sequence-178575-        5435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with thl(CA_C2873) 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 5435)
  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 5435)
  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
FEATURES             Location/Qualifiers
     source          1..5435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1173..1189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1173..1189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1186..1208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1197..1213
                     /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(1221..1227,1228..1245,1246..1251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1266..1287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1404..1421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1575..2163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1655..1674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2334..3125,3126..3194))
                     /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     2957..2976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3195..3299)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3367..3385
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3423..3445)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3545..3564
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3758..3780
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3772..3789
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3773..3789
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaattatcga ttcacaaaaa ataggtacac gaaaaacaag ttaagggatg cagtttatgc
       61 atcccttaac ttacttatta aataatttat agctattgaa aagagataag aattgttcaa
      121 agctaatatt gtttaaatcg tcaattcctg catgttttaa ggaattgtta aattgatttt
      181 ttgtaaatat tttcttgtat tctttgttaa cccatttcat aacgaaataa ttatacttct
      241 gtttatcttt gtgtgatatt cttgattttt ttctatttaa tctgataagt gagctattca
      301 ctttaggttt aggatgaaaa tattctcttg gaaccatact taatatagaa atatcaactt
      361 ctgccattaa aaataatgcc aatgagcgtt ttgtatttaa taatctttta gcaaacccgt
      421 attccacgat taaataaatc tcatcagcta tactatcaaa aacaattttg cgtattatat
      481 ccgtacttat gttataaggt atattaccaa atattttata ggattggttt ttaggaaatt
      541 taaactgcaa tatatccttg tttaaaactt ggaaattatc gtgatcaaca agtttatttt
      601 ctgtagtttt gcataattta tggtctattt caatggcagt tacgaaatta cacctctgta
      661 ctaattcaag ggtaaaatgc ccttttcctg agccgatttc aaagatatta tcatgttcat
      721 ttaatcttat atttgtcatt attttatcta tattatgttt tgaagtaata aagttttgac
      781 tgtgttttat atttttctcg ttcattataa ccctctttat tttttcctcc ttataaaatt
      841 agtataatta tagcacgagc tctgataaat atgaacatga tgagtgatcg ttaaatttat
      901 actgcaatct gatgcgatta ttgaataaaa gatatgagag atttatctag tttctttttt
      961 tacaagaaaa aagaaagttc ttaaaggttt tatacttttg gtcgtagagc acacggttta
     1021 acgacttaat tacgaagtaa ataagtctag tgtgttagac tttaatgttt ttttaaggca
     1081 ttagtgcatt taagcgtcag agcatggctt tatgccgaga aaactattgg ttggaatggc
     1141 gtgtgtgtta gccaaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1201 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1261 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1321 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1381 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1441 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1501 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1741 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1981 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2041 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2221 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2281 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2341 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2401 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2461 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2521 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2581 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2641 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2701 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2761 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2821 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2881 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2941 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3001 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3061 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3121 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3181 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3241 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3301 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3361 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3421 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3481 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3541 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     3601 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     3661 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3721 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3781 acggccagtg aattcccggg gatcctctaa ctaacctcct aaattttgat acggggtaac
     3841 agataaacca tttcaatcta tttcataagt tccatagttt atccctaatt tatacgtttt
     3901 ctctaacaac ttaattatac ccactattat tatttttatc aatatatttt gttaaaaact
     3961 gcagcacatt aagtatatac tattattaaa ctattctata tacttaattt attttaatag
     4021 aaaaacataa tatcataata acttcaaaat taaactttat ttatgatttc atacttgact
     4081 ttgattttag aaaggatata ctttttagca gatttggaaa cggctttgga cgtagtttgc
     4141 ccatagatga acaaacaaac tacatccaaa aattatactt ttcccttcat tggtatccgt
     4201 atttttacat cttaatagcg tatgtattac aacacaccta aacaacgacc ttacggtctg
     4261 ctactgcata tcctagcttg attgtttagt tgcctcaact atgcttaacc ctaccccgaa
     4321 ctcttttttt attgtgggtt ttcgtcgtga agtcccaccg acacataatc ataacataag
     4381 atgtattatg aaaatgcgag tgactatcct tttgtatcgg ctcactacac cacagatata
     4441 ttttttagtg catactgtgt cggcactctc aatattaaat taataaataa attatttttt
     4501 ctttttactc ttctttacat gagctttttt aaagctcctt gcataatatt taatgcatgt
     4561 acgttctttt ttctgttctt cctctgtaaa acatctcatt tttatggcac accatccata
     4621 tcggttcatc tttgaacaat taatacattg gactttccct ttatgtaaac atcttgactc
     4681 attgtattta ctgcaatagg ttgctactgt tttatcatga atcatcaacg aatgtatttt
     4741 gcatatttca gtatctttaa taaactcttt gcaattttta caagttttca tatacgccct
     4801 tcttttcata aattaattta tgaattctat gtattccaag gagcttttta aagctccccc
     4861 tttcgtacta cttagctaca agcactataa aagtcataat gtttactgct aaagtcaata
     4921 atgatattgc taataccttg ttatttgata agatactgct ttcctctgtc actttgctca
     4981 ccccctttca ttttcataaa ttaattatga aaaaataaat atacttctaa tttttatcaa
     5041 ataaaaaagc ctttgcgtac tgcttccaat acacaaaggc tataaacttc taaatcttac
     5101 ttattgcaat ttacatttat atctgttaag ataatctcat aaattgaata tatatagtta
     5161 atgtttctct tgtatgtcgg tacatttgaa atattgctat agatagagtt ctctaacggc
     5221 ttgatgtgtt ggtagcacat ttaagttttg gcttatatac taagttggta gcttaatata
     5281 taagagctga ggacttattt ttttattaaa tttttcaact tgtctatatt ttaacccgta
     5341 attgaataca taacaagtat ttttttttgt attcaattaa acattcataa atgagtataa
     5401 ttaatcatac taaattctat aattttcttt tctgt
//
