LOCUS       sequence-260608-        9203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid with pLL1270 as backbone and 
            alpha-Xylp(Ga0256695_ 1211).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9203)
  TITLE     Expression of arabinoxylan debranching enzymes in 
            Thermoanaerobacterium thermosaccharolyticum- Bacterial expression 
            plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9203)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLL1271
FEATURES             Location/Qualifiers
     source          1..9203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(48..636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(128..147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(807..1598,1599..1667))
                     /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     1430..1449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1668..1772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1840..1858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1896..1918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2018..2037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2231..2253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2245..2262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2246..2262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             3092..3100
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(4822..5616)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     terminator      8898..8925
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(8943..8959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8943..8959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8956..8978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8967..8983
                     /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(8991..8997,8998..9015,9016..9021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9036..9057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
       61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     1861 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     1921 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     1981 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2041 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2101 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2161 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     2221 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgac aaagttttct
     2281 atttgtgtta acattgttta tataatagtg aacagtgtta agattaaatg tgaggtgttt
     2341 gtatggatat taatgattat aaagagaagg gactttattt attaagtagt atggatgatt
     2401 ttattaaaat taatgatttg tttatgggta aagttgtttc tcctggctat gttgcttcgg
     2461 tttttggtgt ttccaggtct actgttacac aatggattca aagacgtaaa attagagctt
     2521 ttaagtataa aggtaaggaa ggtgactata tggttatacc tattgctgat attattgatt
     2581 acaaaagatt gagtaataat gattttattt atgataagtt agtgaggtga tttattttat
     2641 gtttgacgat agctatgttg ttaatgagtg ttcgtctaat gttagtgaaa atgatagaga
     2701 tttttgtagt ttggttggtc gttttatgat tattaatggt atagataagt tggttattaa
     2761 gattaataga aaatttaata ggaaatcttt aagtttagat tttagtgttg atttattccc
     2821 ttctatcaaa gtttctgaat tagttttttt tgatgagttt aacaaaacgt gtggttttta
     2881 tttttctttt aattctttta caatttttaa ggcttttaga gatgttcata atcataataa
     2941 aatatcattt tattttgcat aatttcgggt ctgggccgca gaccaggccc agtgctaaca
     3001 atattaattt ttaatgttag gaattgttta attcttaatt gtgtttttaa aggtagaata
     3061 attacccatt cgccctttag ccaacaaaaa ttaaggaggt ataaacatgg ataaaatgga
     3121 tttgattctt caagatgaaa gactgggtga gatatttaaa gatatagatt taacagataa
     3181 tgaaaagaga tatcttaaat ggttatggaa atgggattat gaaacacgtg atacttttgt
     3241 atcaattttt ttgaagctaa aaaatggtgg aaaatgattt ttttcttatc ttgatatatt
     3301 agaaaaaagc gtactcacga agtaagaatt tgtaaaaaaa gaagggggga tttttttgga
     3361 tgagagtttg tacaagcaga ttttaagtaa tattattatt actcgtgatt attgtaaaaa
     3421 tgttttagat aatataaagt tcaatgaaaa aataattgat tattatgtta tgttacaaaa
     3481 tgatgttttt attgatttta ctaataaaat aaattcaata agggattgta ataaatattg
     3541 gtatttggat gtttataaaa agcagaaaat aaaggatttt aaaaagacta atttgtgtaa
     3601 agataagttc tgtaataatt gtaagaaagt taaacaggct tcaagaatgc aaaaatatat
     3661 tcctgaatta cagaaataca aagatggctt atatcatttt atatttactg ttgaaaatgt
     3721 gccaggtagt gaattaagag atactattga taggttgttt aagtctttta agtcatttac
     3781 aaggtattta agtggtaatc ttaaaataaa aggtgttaat tttgataaat ggggttataa
     3841 aggctgtgta aggtctttag aggtaactta tagtatgatt gataatcata ttatgtatca
     3901 tccacacttg catgttgcga tgatattaga tcctttttac gatggtttta atgttgaaag
     3961 gatgcatata attaataagt ttagttatag ctatggtgtt ttaaaaaggt tgtttactga
     4021 tgatgaatta ttaattcaaa aaatttggta tttattgttt aataatattg aggttaacat
     4081 ggccaatata aataatttag aggatggtta ttcttgttta gttaataagt ttagtgatta
     4141 tgattatgcg gagctgttta agtatatttg taaaaatact gatgaacaag gtttacttat
     4201 gacttatgat atttttaaag atttatattt tgcattacat aatgttcatc agatacaagg
     4261 ctatggttgt ttatataata taagagatga tactcaatta gatttaaagg ttgatgacat
     4321 ttataatgat ttgattgatt tattacaagt tacagaaaat cctatacagt ctatggaaac
     4381 tgtacaggat ttattaaagg atactgaata tacaataata agccgtaagc gtatatttaa
     4441 gtatctaaca caattatatc ataaggattg atatttatac cgtctgtcgg actcatgcgg
     4501 agggggactt gagggggtct cccctcgcat tgtacgacag acggtattat tattatacaa
     4561 atttttttta tgtaattttt tttgtgtaat ttttttatac aaataatatt tcaattcgag
     4621 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcacgataca aattcctcgt
     4681 aggcgctcgg gacccctatc tagcgaactt ttagaaaaga tataaaacat cagagtatgg
     4741 acagttgcgg atgtacttca gaaaagatta gatgtctaaa aagcttgtag ttaaagcttt
     4801 ttagacatct aaatctaggt gctaaaacaa ttcatccagt aaaatataat attttatttt
     4861 ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc
     4921 gatatcctcc ctgatcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc
     4981 gcttctccca agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc
     5041 tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc
     5101 atacagctcg cgcggatctt taaatggagt gtcttcttcc cagttttcgc aatccacatc
     5161 ggccagatcg ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt
     5221 atagggacaa tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc
     5281 gataatcttt tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc
     5341 ctcactcatg agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac
     5401 aggcagcttt ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt
     5461 ccctttatac cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt
     5521 atatacctta gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag
     5581 ttttttcaat tccggtgata ttctcatttt agccatttat tatttccttc cccttttcta
     5641 cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc
     5701 cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg
     5761 gcgtataaca tagtatcgac ggagccgatt ttgaaaccac aattatgata gaatttacaa
     5821 gctataaggt tattgtcctg ggtttcaagc attagtccat gcaagttttt atgctttgcc
     5881 cattctatag atatattgat aagcgcgctg cctatgcctt gccccctgaa atccttacat
     5941 acggcgatat cttctatata aaagatatat tatcttatca gtattgtcaa tatattcaag
     6001 gcaatctgcc tcctcatcct cttcatcctc ttcgtcttgg tagcttttta aatatggcgc
     6061 ttcatagagt aattctgtaa aggtccaatt ctcgttttca tacctcggta taatcttacc
     6121 tatcacctca aatggttcgc tgggtttgag tcgtgactaa gaacgtcaaa gtaattaaca
     6181 atacagctat ttttctcatg cttttacccc tttcataaaa tttaatttta tcgttatcat
     6241 aaaaaattat agacgttata ttgcttgccg ggatatagtg ctgggcattc gttggtgcaa
     6301 aatgttcgga gtaaggtgga tattgatttg catattgatc tattgcattg aaatgattag
     6361 ttatccgtaa atattaatta atcatatcat aaattaatta tatcataatt gttttgacga
     6421 atgaaggttt ttggataaat tatcaagtaa aggaacgcta aaaattttgg cgtaaaatat
     6481 caaaatgacc acttgaatta atatggtaaa gtagatataa tattttggta aacatgcctt
     6541 cagcaaggtt agattagctg tttccgtata aattaaccgt atggtaaaac ggcagtcaga
     6601 aaaataagtc ataagattcc gttatgaaaa tatacttcgg tagttaataa taagagatat
     6661 gaggtgagag atacaagata agagatataa ggtacgaatg tataagatgg tgcttttagg
     6721 cacactaaat aaaaaacaaa taaacgaaaa ttttaaggag gacgaaagat ggcaaagata
     6781 acgattctgg aagacaagat aatttttgaa agaagagatg agcttacggt aatagaagct
     6841 tatggagcaa attgtctgcg ttaccgttca acccgcaata ttaagctgtc tgatgagaac
     6901 tggaccctgc tgccgccgac tacaaaagct gattgtaaaa cctttggaaa tgaaagagaa
     6961 gcaacgatag taaacggcga tatttccgca actataaggg acggctttgg ctggaccgga
     7021 gccagtataa gctactttcg caaggggaag cagattcttc gatcccgtga tgagggtgat
     7081 ccggttaccc gctttaccca cctgggcggt aacaattatc aggtgaaggc gatatttgac
     7141 agtaatccgg gcgagcatat atatggcctg ggacaggagc aggaggatca gtttgaccgc
     7201 aagggctgca cctgcaatat catgcattac aataccaagt cgactttgcc ggttatttat
     7261 tcgtctctgg gatatggctt tttgtggaac aatcctgcac ccggccgttg tgagactacc
     7321 tttaatcata caatgtgggt tgcggatagt gcatatcaga ttgattatct tgtatatgtc
     7381 ggggatactc cggctgacat aatgaaaata tactgtgatc tgacaggcta tgccccgaaa
     7441 ttccctgact gggcggccgg cttctggcag tcaaagctcc gttatgaaag ccaggaggat
     7501 ttgctggagg tggcaaggga atacaagaaa agaggaattc ctctcgcagc gatagtaata
     7561 gattatttcc actggacgga gcagggggaa tggaaatttg atcccaattt gtggcctgat
     7621 cccaaggcaa tgtgtgacga gcttgagtca atgggagtaa agcccgttgt atccatctgg
     7681 cctaccatta atcccaagag tgagaattat catgcaatgg ataatgagaa tatgctggtg
     7741 aggaccgaga acgggcagta tggcatattc aacttttacg gcttacagac ctttatagat
     7801 cccaccaatc ctaaaacagg gccctttgta tggagcaagg taagggagaa ttattataag
     7861 tacggaatta agaacttttg gctggatgag gctgagccgg aggtgcatcc tcagcagttt
     7921 ggcaatctgc gtttttatat aggtaacgga gcccagacgg cattattgta tccatattat
     7981 tatgcaaaga tattttatga tggcttaaga agtgagggag aagaggatat tataatcctt
     8041 acccgggcag cctatcccgg cagccaaaga tttggttccc tggtatggaa cggagacatc
     8101 ccctccacct tcaaggctct tcgccagagt gtaaccagcg gcctttccat gtcaatgtgc
     8161 ggaatcccat ggtggaacag cgacataggc ggcttccatt ccgcagatat agaaagcgac
     8221 tatttcaggg agcttatagt cagatggttc cagttcggcc tctttagtcc ggttatgcgc
     8281 cttcacgggg ccagaaaccg tacaaaaaac caggttgacc gccatccggg cgtgaaggag
     8341 agaagcggcg gagacaatga aatctggagc ttcggtgaaa gaaattatcc tattcttaag
     8401 aatattatag agcttaggga gaggctgaag ccttatattc tcaaatatat ggatatcgcc
     8461 agcaagaccg gctcaccgat tatgaggccg atgttctatg atttttatga tgacgagatt
     8521 tgctaccgac ttgatgatca gtacatgttc ggagaggata tactctttgc tcccattctg
     8581 gagcagggag tgacacagcg cagtgtctac ctgccggagg gtaaatgggt taatgtaaat
     8641 gataaaaaaa cctacgaggg caggcaaatg ataactgcaa gggcagagct gcatgagttt
     8701 attgccttcg taagaagcgg cagcgaggtg ctggaggtat ttagctaact gggcctttcg
     8761 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     8821 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     8881 caggcatcaa attaagcaga aggccatcct gacggatggc cttttagctt ggcgtaatca
     8941 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     9001 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     9061 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagca gatctgcggc
     9121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     9181 caggaaagaa catgtgagca aaa
//
