LOCUS       sequence-305688-        6631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridial vector, carrying lox511/66 and loxFAS/71 sites separated
            by restriction sites in order to add cargo. Use in combination with 
            pQcre1 to insert cargo into lox sites, inserted with pQadd1F/R..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6631)
  AUTHORS   Cerisy T, Rostain W, Chhun A, Boutard M, Salanoubat M, Tolonen AC
  TITLE     A Targetron-Recombinase System for Large-Scale Genome Engineering of
            Clostridia.
  JOURNAL   mSphere. 2019 Dec 11;4(6). pii: 4/6/e00710-19. doi: 
            10.1128/mSphere.00710-19.
  PUBMED    31826971
REFERENCE   2  (bases 1 to 6631)
  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: "mSphere."; 
            date: "2019-12-11"; volume: "4(6). pii: 4/6/e00710-19. doi"; pages: 
            " 10.1128/mSphere.00710-19"
COMMENT     Sequence Label: pQadd2
FEATURES             Location/Qualifiers
     source          1..6631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      391..979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     880..899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1133..1150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1267..1288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1303..1308,1309..1326,1327..1333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1341..1357
                     /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     1346..1368
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1365..1381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1365..1381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(1412..1445)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1518..1535)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1518..1534)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1527..1549)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1860..2231)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2264..2373
                     /note="incP origin of transfer"
ORIGIN
        1 gaattgacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
       61 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      121 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
      181 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
      241 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
      301 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
      361 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      421 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      481 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      541 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      601 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      661 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      721 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      781 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      841 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      901 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      961 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1021 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1081 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1141 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     1201 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     1261 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     1321 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     1381 ccatgattac gccaagcttg catgcctgca gataacttcg tatagtatac attatacgaa
     1441 cggtagagag actagtgtga tatgccctcg aggagatacc gttcgtatat acctttctat
     1501 acgaagttat agaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1561 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1621 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     1681 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg ggatcggtct
     1741 tgccttgctc gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc ttgatggagc
     1801 gcatggggac gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt
     1861 catggctctg ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc
     1921 tcttcgatct tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg
     1981 tcgtcggtga gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg
     2041 gccagctcgc ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg
     2101 gccagcaggt aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt
     2161 cgttcgtctg gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt
     2221 tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga
     2281 gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg
     2341 ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga
     2401 aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc
     2461 gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagat cccccgaagc
     2521 aaacttaaga gtgtgttgat agtgcagtat cttaaaattt tgtgtataat aggaattgaa
     2581 gttaaattag atgctaaaaa tttgtaatta agaaggaggg attcgtcatg ttggtattcc
     2641 aaatgcgtaa tgtagataaa acatctactg ttttgaaaca gactaaaaac agtgattacg
     2701 cagataaata aatacgttag attaattcct accagtgact aatcttatga ctttttaaac
     2761 agataactaa aattacaaac aaatcgttta acttctgtat ttatttacag atgtaatcac
     2821 ttcaggagta attacatgaa caaaaatata aaatattctc aaaacttttt aacgagtgaa
     2881 aaagtactca accaaataat aaaacaattg aatttaaaag aaaccgatac cgtttacgaa
     2941 attggaacag gtaaagggca tttaacgacg aaactggcta aaataagtaa acaggtaacg
     3001 tctattgaat tagacagtca tctattcaac ttatcgtcag aaaaattaaa actgaacatt
     3061 cgtgtcactt taattcacca agatattcta cagtttcaat tccctaacaa acagaggtat
     3121 aaaattgttg ggagtattcc ttaccattta agcacacaaa ttattaaaaa agtggttttt
     3181 gaaagccatg cgtctgacat ctatctgatt gttgaagaag gattctacaa gcgtaccttg
     3241 gatattcacc gaacactagg gttgctcttg cacactcaag tctcgattca gcaattgctt
     3301 aagctgccag cggaatgctt tcatcctaaa ccaaaagtaa acagtgtctt aataaaactt
     3361 acccgccata ccacagatgt tccagataaa tattggaagc tatatacgta ctttgtttca
     3421 aaatgggtca atcgagaata tcgtcaactg tttactaaaa atcagtttca tcaagcaatg
     3481 aaacacgcca aagtaaacaa tttaagtacc attacttatg agcaagtatt gtctattttt
     3541 aatagttatc tattatttaa cgggaggaaa taattctatg agtcgctttt ttaaatttgg
     3601 aaagttacac gttactaaag ggaatggaga taaattatta gatatactac tgacagcttc
     3661 caagaagcta aagaggtccc tagcgcctac ggggaatttg ggcaaatcaa aagaaaaatt
     3721 ctcccaagaa ttagaaatga gtagatcaaa ttattcacga atagaatcag gaaaatcaga
     3781 tccaaccata aaaacactag aacaaattgc aaagttaact aactcaacgc tagtagtgga
     3841 tttaatccca aatgagccaa cagaaccaga accagaaaca gaatcagaac aagtaacatt
     3901 ggatttagaa atggaagaag aaaaaagcaa tgacttcgtg tgaataatgc acgaaatcgt
     3961 tgcttatttt ttttaaaagc ggtatactag atataacgaa acaacgaact gaatagaaac
     4021 gaaaaaagag ccatgacaca tttataaaat gtttgacgac attttataaa tgcatagccc
     4081 gataagattg ccaaaccaac gcttatcagt tagtcagatg aactcttccc tcgtaagaag
     4141 ttatttaatt aactttgttt gaagacggta tataaccgta ctatcattat atagggaaat
     4201 cagagagttt tcaagtatct aagctactga atttaagaat tgttaagcaa tcaatcggaa
     4261 atcgtttgat tgcttttttt gtattcattt atagaaggtg gagtttgtat gaatcatgat
     4321 gaatgtaaaa cttatataaa aaatagttta ttggagataa gaaaattagc aaatatctat
     4381 acactagaaa cgtttaagaa agagttagaa aagagaaata tctacttaga aacaaaatca
     4441 gataagtatt tttcttcgga gggggaagat tatatatata agttaataga aaataacaaa
     4501 ataatttatt cgattagtgg aaaaaaattg acttataaag gaaaaaaatc tttttcaaaa
     4561 catgcaatat tgaaacagtt gaatgaaaaa gcaaaccaag ttaattaaac aacctatttt
     4621 ataggattta taggaaagga gaacagctga atgaatatcc cttttgttgt agaaactgtg
     4681 cttcatgacg gcttgttaaa gtacaaattt aaaaatagta aaattcgctc aatcactacc
     4741 aagccaggta aaagcaaagg ggctattttt gcgtatcgct caaaatcaag catgattggc
     4801 ggtcgtggtg ttgttctgac ttccgaggaa gcgattcaag aaaatcaaga tacatttaca
     4861 cattggacac ccaacgttta tcgttatgga acgtatgcag acgaaaaccg ttcatacacg
     4921 aaaggacatt ctgaaaacaa tttaagacaa atcaatacct tctttattga ttttgatatt
     4981 cacacggcaa aagaaactat ttcagcaagc gatattttaa caaccgctat tgatttaggt
     5041 tttatgccta ctatgattat caaatctgat aaaggttatc aagcatattt tgttttagaa
     5101 acgccagtct atgtgacttc aaaatcagaa tttaaatctg tcaaagcagc caaaataatt
     5161 tcgcaaaata tccgagaata ttttggaaag tctttgccag ttgatctaac gtgtaatcat
     5221 tttggtattg ctcgcatacc aagaacggac aatgtagaat tttttgatcc taattaccgt
     5281 tattctttca aagaatggca agattggtct ttcaaacaaa cagataataa gggctttact
     5341 cgttcaagtc taacggtttt aagcggtaca gaaggcaaaa aacaagtaga tgaaccctgg
     5401 tttaatctct tattgcacga aacgaaattt tcaggagaaa agggtttaat agggcgtaat
     5461 aacgtcatgt ttaccctctc tttagcctac tttagttcag gctattcaat cgaaacgtgc
     5521 gaatataata tgtttgagtt taataatcga ttagatcaac ccttagaaga aaaagaagta
     5581 atcaaaattg ttagaagtgc ctattcagaa aactatcaag gggctaatag ggaatacatt
     5641 accattcttt gcaaagcttg ggtatcaagt gatttaacca gtaaagattt atttgtccgt
     5701 caagggtggt ttaaattcaa gaaaaaaaga agcgaacgtc aacgtgttca tttgtcagaa
     5761 tggaaagaag atttaatggc ttatattagc gaaaaaagcg atgtatacaa gccttattta
     5821 gtgacgacca aaaaagagat tagagaagtg ctaggcattc ctgaacggac attagataaa
     5881 ttgctgaagg tactgaaggc gaatcaggaa attttcttta agattaaacc aggaagaaat
     5941 ggtggcattc aacttgctag tgttaaatca ttgttgctat cgatcattaa agtaaaaaaa
     6001 gaagaaaaag aaagctatat aaaggcgctg acaaattctt ttgacttaga gcatacattc
     6061 attcaagaga ctttaaacaa gctagcagaa cgccctaaaa cggacacaca actcgatttg
     6121 tttagctatg atacaggctg aaaataaaac ccgcactatg ccattacatt tatatctatg
     6181 atacgtgttt gttttttctt tgctgtttag cgaatgatta gcagaaatat acagagtaag
     6241 attttaatta attattaggg ggagaaggag agagtagccc gaaaactttt agttggcttg
     6301 gactgaacga agtgagggaa aggctactaa aacgtcgagg ggcagtgaga gcgaagcgaa
     6361 cacttgattt tttaattttc tatcttttat aggtcattag agtatactta tttgtcctat
     6421 aaactattta gcagcataat agatttattg aataggtcat ttaagttgag catattagag
     6481 gaggaaaatc ttggagaaat atttgaagaa cccgattaca tggattggat tagttcttgt
     6541 ggttacgtgg tttttaacta aaagtagtga atttttgatt tttggtgtgt gtgtcttgtt
     6601 gttagtattt gctagtcaaa gtgattaaat a
//
