LOCUS       sequence-178574-        8031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeted gene disruption by use of a group II intron (targetron) 
            vector in Clostridium acetobutylicum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8031)
  AUTHORS   Shao L, Hu S, Yang Y, Gu Y, Chen J, Yang Y, Jiang W, Yang S
  TITLE     Targeted gene disruption by use of a group II intron (targetron) 
            vector in Clostridium acetobutylicum.
  JOURNAL   Cell Res. 2007 Nov;17(11):963-5. doi: 10.1038/cr.2007.91.
  PUBMED    17971808
REFERENCE   2  (bases 1 to 8031)
  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"; doi: "10.1038/cr.2007"; 
            journalName: "Cell Res"; date: "2007-11"; volume: "17"; issue: "11";
            pages: "963-5"
COMMENT     Sequence Label: pSY6
FEATURES             Location/Qualifiers
     source          1..8031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(67..153)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2329..2351
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5530..5546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5530..5546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5543..5565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5554..5570
                     /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(5578..5584,5585..5602,5603..5608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5623..5644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5761..5778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5932..6520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6012..6031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6691..7482,7483..7551))
                     /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     7314..7333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7552..7656)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7724..7742
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7780..7802)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7902..7921
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(7979..8006)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc
       61 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      121 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
      181 ggggagaccc cacactacca tcggcggctt ggcgatccac aggacgggtg tggtcgccat
      241 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa
      301 gcggtcggac agtgctccga gaacgggtgc tcgacattca cttgtgttta tgaatcacgt
      361 gacgatgaca atgaaagcat acaacaagag ttttacgttg tttcgctatc attgccattt
      421 cccatttttc tttgccttta agatttttga ccttattgac atggtgaatt tcataggaag
      481 tattttcatc agacgtccca cataattcac aacatttagc ttttaacctg ttttcaagag
      541 tattccgggc atagccatac aatacaggag cttgacttat ctcatccgta aattgataag
      601 gggatttaca ttcactaaaa tttgcaaaat aacggcgctg cttaccttgc tttatctcat
      661 acgggatccc ccacgaacca cttccatctt taaacatgga aatggttttt gaaagtgttc
      721 ccttatgttt ggaggctatc gtttttagac agctgtattc cataagataa gcaaaataat
      781 tgagctggtt aaaattactt gctagaccgt agtaattaca aatcccgcgg agttcagaat
      841 tataaattgt gatgatttct aagtctgttg aacgaataag atatttcctg tgaactggaa
      901 accatgagct atctttcttt tggatagcta ttttcttgtc aaaaataaat tgacgaattt
      961 tgtcttgaag aggaataagg agttctacac tcccattgag tgttctcttt ttgactttac
     1021 cagatcgttt tatcgttcca gatctcctta ctcgtatatc atatcccaga aaacgagcgg
     1081 gttgactgct atgtgtgatg agtgtttttt cttcactcaa ttccattttt agcttgttat
     1141 gaataaaaag ttttaattgt tcttttatcc attgacagtc ctctttgctt cctttaacag
     1201 agataatgaa gtcgtccgca taccggacgt atttcaatac tttatttgtc tgtgaggtac
     1261 aggggagtgt gggtaatctt ttacgttttt cttgatattc taaaagaact ttagcttttt
     1321 cttcaccctc caacttcttg agacggtgag aaattctttt tatctcattg tggagctccc
     1381 gatattcagg tgttattctt tctggacttt ctcggtcaaa cttcattttg agttgtaaaa
     1441 caaacttatc caattcatga agatagatgt tggccaaaag aggagataga attccacctt
     1501 gaggtgttcc gctgtaagtt ttgtgatact gccagttttc cagataacct gcttttagaa
     1561 atttataaat caattggctc attttcatat ctttgatttt aagattgatg agtccaatga
     1621 gtgtaacgtg gtctatatta tcgaagcagc cttttatatc tccctccaca aaccatcttg
     1681 cgccgccaaa ctctcttttg attgttttca aagctgtgtg acagcttcgt tgaggtctaa
     1741 aaccgtgaga cacatcttcg aataccggtt catagataga ttcaagaatt attctcacag
     1801 cttcttggat caatttatct gtgaaagttg gaattcctaa aggtctcatc tttttagaat
     1861 tcttttttgc aatatacatt cttcgtacag gttgaggata gtaagttccg tcttttaaag
     1921 attgaataat cttttttatt ttttcttcac taaagccatc cgctgtatca tctaatattc
     1981 cttttgtgga agctccttta ttggaatata aattttgata cgccacgtaa taaatatctg
     2041 gacgtaaaag ataacgataa agtcttgtaa aaacttcgtc tatattttct tgtgaatttt
     2101 tactgattct ttctaaaatt gccattgttg gtttcatttt gaggttttcc tccctaatca
     2161 atttttaatt ttagtacaca ataactgtac ccctttgcca tgtaaagggc gttacccttc
     2221 tcagactact acgagtgcat agaaattgca tcaacgcata tagcgctagc agcacgccat
     2281 agtgactggc gatgctgtcg gaatggacga tctagccata tatgtatatc tccttcttaa
     2341 agttaaacaa aattatttct agggggctgc agaaaatgat atggtgaagt agggaggtac
     2401 cgccttgttc acattactgt gactggtttg caccaccctc ttcgggaacc gtacgtaccc
     2461 ctctcggagt atacggctct gttattgttc gttcgtaaaa attcactgtc gacattcact
     2521 tgtgtttatg aatcacgtga cgatgacaat gaaagcatac aacaagagtt ttacgttgtt
     2581 tcgctatcat tgccatttcc caacgcgtcg ccacgtaata aatatctgga cgtaaaagat
     2641 aacgataaag tcttgtaaaa acttcgtcta tattttcttg tgaattttta ctgattcttt
     2701 ctaaaattgc cattgttggt ttcattttga ggttttcctc cctaatcaat ttttaatttt
     2761 agtacacaat aactgtaccc ctttgccatg taaagggcgt tacccttctc agactactac
     2821 gagtactccg tacccttgca agattttcaa gctctagtgc tatagccttt ttcctccttt
     2881 ctattaggca ttcttgttta gggtatcccc agttagtgtt aagtcttggt aaattcagat
     2941 tctcggcatc gctttcgttt cgttcccata ggttctccta cagattgtac aaatgtggtg
     3001 ataacagata agtccatagg cttaacttac ctttctttgt actagaggtt tcagacactt
     3061 tcctctatcg accagcaacc gaaattagaa acttgcgttc agtaaacaca acttatacct
     3121 tatatctgat taacattgcg actcagtcgt acccgattgt ctttaggtaa ctcatcgctt
     3181 tccaaccgtg ctctgttccc gtatcagctt tcgcttttcg gttaggttgg ctgttttctg
     3241 tgttatctta cagagtagta ccttaaacta cttgacttaa caccctatct gggcgcaccc
     3301 tatgggctgg taaggataat tatctcgagc atatgtcgac actccctttt actatttaat
     3361 tatctatgtt aaatgattaa cgtgctcatt tatattttaa caaacttttc acatgaagtt
     3421 aagtttttta gaaaattatt tatattttat agaattcccg gggatccgtc gacctgcagc
     3481 caagcttaat cgcatttcat agattgacct cccaataact acgtggtgtt attgggaggt
     3541 caatctattt catttgcctc ttgctcaaag ttcccaaatt cgagtaagag gtatttttgt
     3601 ttttggtcgt cgcctctcat tagtagttca gggtttaaca ttaatactcc agtttttctt
     3661 tttataatat ttccttcttc taagatttta agtgttgtta ttactgtttg tagacttgtt
     3721 cctgtagctt ttgctatttc tcttgttgta gctatcattg tattgttact taagtggaca
     3781 ttatctagga tatagttaac gattttaagt ttttttccgc caatcatatc taacatactt
     3841 attaattgca ctatatatgc ctttacgaag ttaccagacg tttgtttacg gtataacttg
     3901 tctacctcta tgacttctcc actttcttcg tctatgagcc tctgagagcc tttatagact
     3961 gttccatatc tttctttcat ctttttctca ctccttattt taaactattc taactatatc
     4021 ataactgttc taaaaaaaaa agaacatttg ttaaaagaaa ttagaacaaa atgagtgaaa
     4081 aattagaaca aacaaattcc ttataaacct tatcatctca acctatatta agattttacc
     4141 tagttgaatc ttcttttcta tataaagcgt cggagcatat cagggggtta tctaacgtaa
     4201 atgctaccct tcggctcgct ttcgctcggc attgacgtca gatactgcac cccctgaacc
     4261 cccatgctcc aacagcaaaa aggaaacttt ttgctgcttt tccgacgctt attcgcttcg
     4321 ctcatattta tatagaaaag aagtgaatgc gcaaaagaca taatcgattc acaaaaaata
     4381 ggtacacgaa aaacaagtta agggatgcag tttatgcatc ccttaactta cttattaaat
     4441 aatttatagc tattgaaaag agataagaat tgttcaaagc taatattgtt taaatcgtca
     4501 attcctgcat gttttaagga attgttaaat tgattttttg taaatatttt cttgtattct
     4561 ttgttaaccc atttcataac gaaataatta tacttctgtt tatctttgtg tgatattctt
     4621 gatttttttc tatttaatct gataagtgag ctattcactt taggtttagg atgaaaatat
     4681 tctcttggaa ccatacttaa tatagaaata tcaacttctg ccattaaaaa taatgccaat
     4741 gagcgttttg tatttaataa tcttttagca aacccgtatt ccacgattaa ataaatctca
     4801 tcagctatac tatcaaaaac aattttgcgt attatatccg tacttatgtt ataaggtata
     4861 ttaccaaata ttttatagga ttggttttta ggaaatttaa actgcaatat atccttgttt
     4921 aaaacttgga aattatcgtg atcaacaagt ttattttctg tagttttgca taatttatgg
     4981 tctatttcaa tggcagttac gaaattacac ctctgtacta attcaagggt aaaatgccct
     5041 tttcctgagc cgatttcaaa gatattatca tgttcattta atcttatatt tgtcattatt
     5101 ttatctatat tatgttttga agtaataaag ttttgactgt gttttatatt tttctcgttc
     5161 attataaccc tctttatttt ttcctcctta taaaattagt ataattatag cacgagctct
     5221 gataaatatg aacatgatga gtgatcgtta aatttatact gcaatctgat gcgattattg
     5281 aataaaagat atgagagatt tatctagttt ctttttttac aagaaaaaag aaagttctta
     5341 aaggttttat acttttggtc gtagagcaca cggtttaacg acttaattac gaagtaaata
     5401 agtctagtgt gttagacttt aatgtttttt taaggcatta gtgcatttaa gcgtcagagc
     5461 atggctttat gccgagaaaa ctattggttg gaatggcgtg tgtgttagcc aaagcttggc
     5521 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5581 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5641 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5701 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5761 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5821 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6661 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6721 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6781 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6841 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6901 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6961 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7021 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7081 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7141 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7201 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7261 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7321 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7381 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7441 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7501 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7561 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7621 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7681 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7741 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7801 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7861 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7921 actgagagtg caccatatgc ttgtagatat gacgacagga agagtttgta gaaacgcaaa
     7981 aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt a
//
