LOCUS       sequence-260567-        7223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector used for enzyme expression in Thermoanaerobacterium. 
            thermosaccharolyticum throughout this study. Contains the CBP 
            promoter from Clostridium thermocellum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7223)
  TITLE     Expression of arabinoxylan debranching enzymes in 
            Thermoanaerobacterium thermosaccharolyticum- Bacterial expression 
            plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7223)
  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            "Unpublished"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: pLL1270
FEATURES             Location/Qualifiers
     source          1..7223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..24
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8..24
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(42..69)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             1371..2165
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     RBS             3887..3895
                     /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)"
     primer_bind     complement(4725..4742)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4725..4741)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4734..4756)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4950..4969)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5069..5091
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5129..5147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5215..5319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5320..5388,5389..6180)
                     /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(5538..5557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6351..6939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6840..6859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7133..7154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7169..7174,7175..7192,7193..7199)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7207..7223
                     /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     join(7212..7223,1..11)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 tttcacacag gaaacagcta tgaccatgat tacgccaagc taaaaggcca tccgtcagga
       61 tggccttctg cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
      121 cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
      181 cgttcaccga caaacaacag ataaaacgaa aggcccagct ttcgtcctcc ttaaaatttt
      241 cgtttatttg ttttttattt agtgtgccta aaagcaccat cttatacatt cgtaccttat
      301 atctcttatc ttgtatctct cacctcatat ctcttattat taactaccga agtatatttt
      361 cataacggaa tcttatgact tatttttctg actgccgttt taccatacgg ttaatttata
      421 cggaaacagc taatctaacc ttgctgaagg catgtttacc aaaatattat atctacttta
      481 ccatattaat tcaagtggtc attttgatat tttacgccaa aatttttagc gttcctttac
      541 ttgataattt atccaaaaac cttcattcgt caaaacaatt atgatataat taatttatga
      601 tatgattaat taatatttac ggataactaa tcatttcaat gcaatagatc aatatgcaaa
      661 tcaatatcca ccttactccg aacattttgc accaacgaat gcccagcact atatcccggc
      721 aagcaatata acgtctataa ttttttatga taacgataaa attaaatttt atgaaagggg
      781 taaaagcatg agaaaaatag ctgtattgtt aattactttg acgttcttag tcacgactca
      841 aacccagcga accatttgag gtgataggta agattatacc gaggtatgaa aacgagaatt
      901 ggacctttac agaattactc tatgaagcgc catatttaaa aagctaccaa gacgaagagg
      961 atgaagagga tgaggaggca gattgccttg aatatattga caatactgat aagataatat
     1021 atcttttata tagaagatat cgccgtatgt aaggatttca gggggcaagg cataggcagc
     1081 gcgcttatca atatatctat agaatgggca aagcataaaa acttgcatgg actaatgctt
     1141 gaaacccagg acaataacct tatagcttgt aaattctatc ataattgtgg tttcaaaatc
     1201 ggctccgtcg atactatgtt atacgccaac tttgaaaaca actttgaaaa agctgttttc
     1261 tggtatttaa ggttttagaa tgcaaggaac agtgaattgg agttcgtctt gttataatta
     1321 gcttcttggg gtatctttaa atactgtaga aaaggggaag gaaataataa atggctaaaa
     1381 tgagaatatc accggaattg aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg
     1441 aaggaatgtc tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa
     1501 aaatgacgga cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga
     1561 tgctatggct ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg
     1621 gctggagcaa tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag
     1681 atgaacaaag ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact
     1741 ccatcgacat atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg
     1801 attacttact gaataacgat ctggccgatg tggattgcga aaactgggaa gaagacactc
     1861 catttaaaga tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac
     1921 ttgtcttttc ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa
     1981 gtggctttat tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct
     2041 gcgtccggtc gatcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact
     2101 tactggggat caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt
     2161 tttagcacct agatttagat gtctaaaaag ctttaactac aagcttttta gacatctaat
     2221 cttttctgaa gtacatccgc aactgtccat actctgatgt tttatatctt ttctaaaagt
     2281 tcgctagata ggggtcccga gcgcctacga ggaatttgta tcgtgcatgc ctgcaggtcg
     2341 actctagagg atccccgggt accgagctcg aattgaaata ttatttgtat aaaaaaatta
     2401 cacaaaaaaa attacataaa aaaaatttgt ataataataa taccgtctgt cgtacaatgc
     2461 gaggggagac cccctcaagt ccccctccgc atgagtccga cagacggtat aaatatcaat
     2521 ccttatgata taattgtgtt agatacttaa atatacgctt acggcttatt attgtatatt
     2581 cagtatcctt taataaatcc tgtacagttt ccatagactg tataggattt tctgtaactt
     2641 gtaataaatc aatcaaatca ttataaatgt catcaacctt taaatctaat tgagtatcat
     2701 ctcttatatt atataaacaa ccatagcctt gtatctgatg aacattatgt aatgcaaaat
     2761 ataaatcttt aaaaatatca taagtcataa gtaaaccttg ttcatcagta tttttacaaa
     2821 tatacttaaa cagctccgca taatcataat cactaaactt attaactaaa caagaataac
     2881 catcctctaa attatttata ttggccatgt taacctcaat attattaaac aataaatacc
     2941 aaattttttg aattaataat tcatcatcag taaacaacct ttttaaaaca ccatagctat
     3001 aactaaactt attaattata tgcatccttt caacattaaa accatcgtaa aaaggatcta
     3061 atatcatcgc aacatgcaag tgtggatgat acataatatg attatcaatc atactataag
     3121 ttacctctaa agaccttaca cagcctttat aaccccattt atcaaaatta acacctttta
     3181 ttttaagatt accacttaaa taccttgtaa atgacttaaa agacttaaac aacctatcaa
     3241 tagtatctct taattcacta cctggcacat tttcaacagt aaatataaaa tgatataagc
     3301 catctttgta tttctgtaat tcaggaatat atttttgcat tcttgaagcc tgtttaactt
     3361 tcttacaatt attacagaac ttatctttac acaaattagt ctttttaaaa tcctttattt
     3421 tctgcttttt ataaacatcc aaataccaat atttattaca atcccttatt gaatttattt
     3481 tattagtaaa atcaataaaa acatcatttt gtaacataac ataataatca attatttttt
     3541 cattgaactt tatattatct aaaacatttt tacaataatc acgagtaata ataatattac
     3601 ttaaaatctg cttgtacaaa ctctcatcca aaaaaatccc cccttctttt tttacaaatt
     3661 cttacttcgt gagtacgctt ttttctaata tatcaagata agaaaaaaat cattttccac
     3721 cattttttag cttcaaaaaa attgatacaa aagtatcacg tgtttcataa tcccatttcc
     3781 ataaccattt aagatatctc ttttcattat ctgttaaatc tatatcttta aatatctcac
     3841 ccagtctttc atcttgaaga atcaaatcca ttttatccat gtttatacct ccttaatttt
     3901 tgttggctaa agggcgaatg ggtaattatt ctacctttaa aaacacaatt aagaattaaa
     3961 caattcctaa cattaaaaat taatattgtt agcactgggc ctggtctgcg gcccagaccc
     4021 gaaattatgc aaaataaaat gatattttat tatgattatg aacatctcta aaagccttaa
     4081 aaattgtaaa agaattaaaa gaaaaataaa aaccacacgt tttgttaaac tcatcaaaaa
     4141 aaactaattc agaaactttg atagaaggga ataaatcaac actaaaatct aaacttaaag
     4201 atttcctatt aaattttcta ttaatcttaa taaccaactt atctatacca ttaataatca
     4261 taaaacgacc aaccaaacta caaaaatctc tatcattttc actaacatta gacgaacact
     4321 cattaacaac atagctatcg tcaaacataa aataaatcac ctcactaact tatcataaat
     4381 aaaatcatta ttactcaatc ttttgtaatc aataatatca gcaataggta taaccatata
     4441 gtcaccttcc ttacctttat acttaaaagc tctaatttta cgtctttgaa tccattgtgt
     4501 aacagtagac ctggaaacac caaaaaccga agcaacatag ccaggagaaa caactttacc
     4561 cataaacaaa tcattaattt taataaaatc atccatacta cttaataaat aaagtccctt
     4621 ctctttataa tcattaatat ccatacaaac acctcacatt taatcttaac actgttcact
     4681 attatataaa caatgttaac acaaatagaa aactttgtca attcactggc cgtcgtttta
     4741 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4801 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4861 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4921 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4981 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5041 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5101 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5521 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6481 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagatct
     7081 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7141 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7201 gtggaattgt gagcggataa caa
//
