LOCUS       sequence-357165-        6356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TtEnd_CDC_pEHISTEV expresses tuncated variant of TtEnd5A (186-531 
            amni caids) comprsisng of cataytic domain and a linker at 
            C-terminal. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6356)
  AUTHORS   Hussain N, Mikolajek H, Harrison PJ, Paterson N, Akhtar MW, Sadaf S,
            Naismith JH
  TITLE     Structural and functional snapshots of a broad-specificity 
            endoglucanase from Thermogutta terrifontis for biomass 
            saccharification.
  JOURNAL   Arch Biochem Biophys. 2025 Feb;764:110274. doi: 
            10.1016/j.abb.2024.110274. Epub 2024 Dec 17.
  PUBMED    39701201
REFERENCE   2  (bases 1 to 6356)
  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: "Arch 
            Biochem Biophys."; date: "2025-02"; volume: "764:110274. doi"; 
            pages: " 10.1016/j.abb.2024.110274. Epub 2024 Dec 17"
COMMENT     Sequence Label: TtEnd_CDC_pEHISTEV
FEATURES             Location/Qualifiers
     source          1..6356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      59..514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(146..165)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     356..377
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(606..1421)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(606..1421)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1329..1348
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1413..1432
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1543..2131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2032..2051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2285..2302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2317..2456
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2382..2401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2542..2564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2558..2749)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3558..4640)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4602..4621
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4641..4718)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4924..4943
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5031..5050
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5031..5049
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5050..5074
                     /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)."
     RBS             5089..5111
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5131..5148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5170..5190
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             6247..6264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6317..6335)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(6331..6356,1..22)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
       61 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      121 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      181 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      241 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      301 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      361 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      421 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      481 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
      541 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
      601 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
      661 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      721 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
      781 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
      841 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
      901 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
      961 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1021 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1081 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1141 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1201 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1261 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     1321 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1381 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     1441 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     1501 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1561 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1621 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1681 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1741 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1801 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1861 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1921 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1981 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2041 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2101 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2161 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2221 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2281 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2341 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     2401 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2461 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2521 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2581 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2641 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2701 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2761 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2821 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2881 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2941 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     3001 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     3061 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3121 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3181 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     3241 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     3301 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     3361 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     3421 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     3481 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     3541 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3601 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     3661 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3721 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     3781 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     3841 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     3901 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     3961 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4021 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4081 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4141 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4201 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4261 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4321 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4381 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4441 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4501 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     4561 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     4621 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     4681 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     4741 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     4801 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     4861 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     4921 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     4981 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact
     5041 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     5101 taagaaggag atatacatat gtcgtactac catcaccatc accatcacga ttacgatatc
     5161 ccaacgaccg aaaacctgta ttttcagggc gccatggctc tgagcagcaa ccataaagaa
     5221 gtgtttaaag gtcataatct gcctcgtctg cgtggtgcaa tgattggtcc gcatgttacc
     5281 aatgccgatc tgctggaatt tggtaatgtt tggaaagcca atcatattcg ctggcagctg
     5341 atttggaatg gttttccgca tagtccggca gatagcgcaa ccctggatga atatcgtcag
     5401 tggctggatg gtgcactgaa acgtctggaa gcagcactgc cggtttgtcg tgaagcaggt
     5461 attctggtta ccgtggatct gcatacaccg cctggtggcc gtaatgaagc aagcgaatgt
     5521 cgtatttttc acgatcgcga atttcagaaa gcctttattg acatctggga agatattgca
     5581 cgtcgttttg cagatagtga tgttgtttgg ggttatgatc tggttaatga accggttgaa
     5641 ggtatggtgc cggatggtct gatgaattgg cagcgtctgg ccgaagaaac cgcacgtcgt
     5701 gtgcgtgcaa ttgatcagaa acatgcaatt atcattgaac cggcaccgtg gggtagcccg
     5761 agcagcattg ctctgctgga tccgattgat gttccgggtg ttgtttatag cgttcatatg
     5821 tatgttccgc atgcatttac ccatcagggt gtttatgata atccggtggg tattgtttat
     5881 ccgggtacaa ttgatggcaa atggtatgat cgtaataccc tgcgtaaagt tctggaaccg
     5941 gtgcgtcgtt ttcaagaaga aaacggggtt catatctata tcggtgaatt tagcgcaatt
     6001 cgttgggcac cagccgatag cgcatgtcag tatctgaaag attgcatcga aatcttcgag
     6061 gaatatggtt gggattgggc atatcatgca tttcgtgaat gggatggttg gagcgttgaa
     6121 catggtccgg atcgtaatga tcgcaatcgt accgcaacac cgaccgatcg tgccctgctg
     6181 ctgcgtagct ggtatgcaga aaatgtgaaa ccgcagttta gcgacaaaaa gaaggattaa
     6241 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     6301 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaa
//
