LOCUS       sequence-178582-        4865 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 4865)
  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 4865)
  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: pIMP1-Pthl
FEATURES             Location/Qualifiers
     source          1..4865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     309..326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    443..464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(479..484,485..502,503..509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    517..533
                     /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     522..544
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     541..557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     541..557
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2806..2823)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2806..2822)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2815..2837)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3031..3050)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3150..3172
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3210..3228)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3296..3400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3401..3469,3470..4261)
                     /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(3619..3638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4432..4865,1..155)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
       61 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      121 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      181 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      241 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      301 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      421 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      481 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      541 caggaaacag ctatgaccat gattacgcca agctttggct aacacacacg ccattccaac
      601 caatagtttt ctcggcataa agccatgctc tgacgcttaa atgcactaat gccttaaaaa
      661 aacattaaag tctaacacac tagacttatt tacttcgtaa ttaagtcgtt aaaccgtgtg
      721 ctctacgacc aaaagtataa aacctttaag aactttcttt tttcttgtaa aaaaagaaac
      781 tagataaatc tctcatatct tttattcaat aatcgcatca gattgcagta taaatttaac
      841 gatcactcat catgttcata tttatcagag ctcgtgctat aattatacta attttataag
      901 gaggaaaaaa taaagagggt tataatgaac gagaaaaata taaaacacag tcaaaacttt
      961 attacttcaa aacataatat agataaaata atgacaaata taagattaaa tgaacatgat
     1021 aatatctttg aaatcggctc aggaaaaggg cattttaccc ttgaattagt acagaggtgt
     1081 aatttcgtaa ctgccattga aatagaccat aaattatgca aaactacaga aaataaactt
     1141 gttgatcacg ataatttcca agttttaaac aaggatatat tgcagtttaa atttcctaaa
     1201 aaccaatcct ataaaatatt tggtaatata ccttataaca taagtacgga tataatacgc
     1261 aaaattgttt ttgatagtat agctgatgag atttatttaa tcgtggaata cgggtttgct
     1321 aaaagattat taaatacaaa acgctcattg gcattatttt taatggcaga agttgatatt
     1381 tctatattaa gtatggttcc aagagaatat tttcatccta aacctaaagt gaatagctca
     1441 cttatcagat taaatagaaa aaaatcaaga atatcacaca aagataaaca gaagtataat
     1501 tatttcgtta tgaaatgggt taacaaagaa tacaagaaaa tatttacaaa aaatcaattt
     1561 aacaattcct taaaacatgc aggaattgac gatttaaaca atattagctt tgaacaattc
     1621 ttatctcttt tcaatagcta taaattattt aataagtaag ttaagggatg cataaactgc
     1681 atcccttaac ttgtttttcg tgtacctatt ttttgtgaat cgattatgtc ttttgcgcat
     1741 tcacttcttt tctatataaa tatgagcgaa gcgaataagc gtcggaaaag cagcaaaaag
     1801 tttccttttt gctgttggag catgggggtt cagggggtgc agtatctgac gtcaatgccg
     1861 agcgaaagcg agccgaaggg tagcatttac gttagataac cccctgatat gctccgacgc
     1921 tttatataga aaagaagatt caactaggta aaatcttaat ataggttgag atgataaggt
     1981 ttataaggaa tttgtttgtt ctaatttttc actcattttg ttctaatttc ttttaacaaa
     2041 tgttcttttt tttttagaac agttatgata tagttagaat agtttaaaat aaggagtgag
     2101 aaaaagatga aagaaagata tggaacagtc tataaaggct ctcagaggct catagacgaa
     2161 gaaagtggag aagtcataga ggtagacaag ttataccgta aacaaacgtc tggtaacttc
     2221 gtaaaggcat atatagtgca attaataagt atgttagata tgattggcgg aaaaaaactt
     2281 aaaatcgtta actatatcct agataatgtc cacttaagta acaatacaat gatagctaca
     2341 acaagagaaa tagcaaaagc tacaggaaca agtctacaaa cagtaataac aacacttaaa
     2401 atcttagaag aaggaaatat tataaaaaga aaaactggag tattaatgtt aaaccctgaa
     2461 ctactaatga gaggcgacga ccaaaaacaa aaatacctct tactcgaatt tgggaacttt
     2521 gagcaagagg caaatgaaat agattgacct cccaataaca ccacgtagtt attgggaggt
     2581 caatctatga aatgcgatta agcttggctg cagtttttaa caaaatatat tgataaaaat
     2641 aataatagtg ggtataatta agttgttaga gaaaacgtat aaattaggga taaactatgg
     2701 aacttatgaa atagattgaa atggtttatc tgttaccccg tatcaaaatt taggaggtta
     2761 gttagagtcg actctagagg atccccgggt accgagctcg aattcactgg ccgtcgtttt
     2821 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2881 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2941 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3001 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3061 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3121 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3181 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3241 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3301 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3361 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3421 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3481 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3541 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3601 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3661 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3721 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3781 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3841 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3901 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3961 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4021 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4081 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4141 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4201 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4261 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4321 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4381 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4441 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4501 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4561 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4801 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4861 ggaca
//
