LOCUS       sequence-225594-        9810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Eno-promoter driven expression vector for protein production in 
            Trichoderma reesei.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9810)
  AUTHORS   Linger JG, Taylor LE 2nd, Baker JO, Vander Wall T, Hobdey SE, 
            Podkaminer K, Himmel ME, Decker SR
  TITLE     A constitutive expression system for glycosyl hydrolase family 7 
            cellobiohydrolases in Hypocrea jecorina.
  JOURNAL   Biotechnol Biofuels. 2015 Mar 18;8:45. doi: 
            10.1186/s13068-015-0230-2. eCollection 2015.
  PUBMED    25904982
REFERENCE   2  (bases 1 to 9810)
  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: "Biotechnol 
            Biofuels."; date: "2015-03-18"; volume: "8:45. doi"; pages: " 
            10.1186/s13068-015-0230-2. eCollection 2015"
COMMENT     Sequence Label: pTrEno
FEATURES             Location/Qualifiers
     source          1..9810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7589..7605)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7589..7605)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7602..7624)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7613..7629
                     /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(7637..7643,7644..7661,7662..7667))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7682..7703
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7820..7837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7991..8579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8071..8090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8750..9541,9542..9610))
                     /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     9373..9392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9611..9715)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9783..9801
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 acgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ggagatcggt acttcgcgaa
      421 tgcgtcgaga cctgcaggtg attccgtcct ggattgccaa ctccttgacg ccaaacggcc
      481 cgatagaatt gccctgcccc tgatcgctcg acagccttgc cggccatcgt aagcataggc
      541 acaagattcg aaccgatctc gcttctccct ctagttctgt cgaggcttga tcatgccctc
      601 ctgcacgatg tagtaaacga cggtagagtc attgtgcagc gcagcgagga caatgcgctt
      661 cgcccgcttg cgagcctccg gaagggcgtc catgacggcg gcgaggctgg acagcgtcca
      721 cttctcggcg agatggacgg ccagcaccca ttcgtactcg gcgtcgtgct gcgcccttgt
      781 ccctgttgac ttttcatatg caagcgctgc tatctggtcg tcggggtgga gatacagaag
      841 gcgtggcggg aggccgcgga tcattggccg cgacaggcct gggaggcgat ggatttgcag
      901 cgaggcccat tggtgctgct cttgcaggtt ctcaaatacc gcagaggcga catcggccgc
      961 gaactgatgt gacgcatcca tggggggggg gaggctcaat ggacgtgtag gtactacgta
     1021 taggcagtac tatgtaggaa gtaagacaaa tgacgagaga atgagaagcg agaagcgagc
     1081 gggatcagaa atggtcgcaa tgatattaca aagcgctgcg tctttcgtcc atgatcttct
     1141 tcgtatctta ggtatctgcc cgcatcaggc caagcggccg ctgcgtcgcg atagaaactt
     1201 tttcgctgag aggccggaga aataggacta gcgtttctcg ctgtctaccg attcacgtaa
     1261 cccggcaggc aagtggccca attattacct cacaagcacc ccagctgcgc ctgccctgcc
     1321 ggttgagagc tccgtttctc gttaggtaac ttgcagtgct aatccagtta tctgcgaggg
     1381 aaggcatgac ttgccccacc cgtctcgctc tctctggacc cctccatttc tctcgacaac
     1441 aaaaacacca tccttcttcc ttttccctcc ccaagattct ctcgacttgc tcaatcacgc
     1501 ctcacagcaa gagcctgaga cactcaatcc atttctccac tacaggtacg gtgcctccct
     1561 ttacttgatc tctacaagag gcagaccttg ttctctgtcg tccaagcttc tgcccctctt
     1621 tcccgtttca gccttcccag ctacccctcc atttgggggg gcagagctgc tgaaagacgc
     1681 ggcgtctggc ggaacgtgtt gaggcatatt ctcaatatac ttgtttcgct cctctggctg
     1741 gattttgcta tccgcgggag ctcttgtata ccagaggaat gatgcattgc catcgaactt
     1801 ggctccgtca tctcgcgcgg gcttcacttt cgatgtcgtg atcgcccagc tacctacctc
     1861 ccgagatagc ttcgaataca cctgctaaca gacattgggt ccatccagcc acctgaaata
     1921 gcttcaaatt aattaaatgt atcggaagtt ggccgtcatc tcggccttct tggccacagc
     1981 tcgtgctcag tcggcctgca ctctccaatc ggagactcac ccgcctctga catggcagaa
     2041 atgctcgtct ggtggcacgt gcactcaaca gacaggctcc gtggtcatcg acgccaactg
     2101 gcgctggact cacgctacga acagcagcac gaactgctac gatggcaaca cttggagctc
     2161 gaccctatgt cctgacaacg agacctgcgc gaagaactgc tgtctggacg gtgccgccta
     2221 cgcgtccacg tacggagtta ccacgagcgg taacagcctc tccattggct ttgtcaccca
     2281 gtctgcgcag aagaacgttg gcgctcgcct ttaccttatg gcgagcgaca cgacctacca
     2341 ggaattcacc ctgcttggca acgagttctc tttcgatgtt gatgtttcgc agctgccgtg
     2401 cggcttgaac ggagctctct acttcgtgtc catggacgcg gatggtggcg tgagcaagta
     2461 tcccaccaac accgctggcg ccaagtacgg cacggggtac tgtgacagcc agtgtccccg
     2521 cgatctgaag ttcatcaatg gccaggccaa cgttgagggc tgggagccgt catccaacaa
     2581 cgcgaacacg ggcattggag gacacggaag ctgctgctct gagatggata tctgggaggc
     2641 caactccatc tccgaggctc ttacccccca cccttgcacg actgtcggcc aggagatctg
     2701 cgagggtgat gggtgcggcg gaacttactc cgataacaga tatggcggca cttgcgatcc
     2761 cgatggctgc gactggaacc cataccgcct gggcaacacc agcttctacg gccctggctc
     2821 aagctttacc ctcgatacca ccaagaaatt gaccgttgtc acccagttcg agacgtcggg
     2881 tgccatcaac cgatactatg tccagaatgg cgtcactttc cagcagccca acgccgagct
     2941 tggtagttac tctggcaacg agctcaacga tgattactgc acagctgagg aggcagaatt
     3001 cggcggatcc tctttctcag acaagggcgg cctgactcag ttcaagaagg ctacctctgg
     3061 cggcatggtt ctggtcatga gtctgtggga tgattactac gccaacatgc tgtggctgga
     3121 ctccacctac ccgacaaacg agacctcctc cacacccggt gccgtgcgcg gaagctgctc
     3181 caccagctcc ggtgtccctg ctcaggtcga atctcagtct cccaacgcca aggtcacctt
     3241 ctccaacatc aagttcggac ccattggcag caccggcaac cctagcggcg gcaaccctcc
     3301 cggcggaaac ccgcctggca ccaccaccac ccgccgccca gccactacca ctggaagctc
     3361 tcccggacct acccagtctc actacggcca gtgcggcggt attggctaca gcggccccac
     3421 ggtctgcgcc agcggcacaa cttgccaggt cctgaaccct tactactctc agtgcctgta
     3481 atctagaggc tttcgtgacc gggcttcaaa caatgatgtg cgatggtgtg gttcccggtt
     3541 ggcggagtct ttgtctactt tggttgtctg tcgcaggtcg gtagaccgca aatgagcaac
     3601 tgatggattg ttgccagcga tactataatt cacatggatg gtctttgtcg atcagtagct
     3661 agtgagagag agagaacatc tatccacaat gtcgagtgtc tattagacat actccgagaa
     3721 taaagtcaac tgtgtctgtg atctaaagat cgattcggca gtcgagtagc gtataacaac
     3781 tccgagtacc agcaaaagca cgtcgtgaca ggagcagggc tttgccaact gcgcaacctt
     3841 gcttgaatga ggatacacgg ggtgcaacat ggctgtactg atccatcgca accaaaattt
     3901 ctgtttatag atcaagctgg tagattccaa ttactccacc tcttgcgctt ctccatgaca
     3961 tgtaagtgca cgtggaaacg cggccgcggt tgagctaaca cgccacgcgt tctgcggcat
     4021 ctagttgatg ccaagatgat ggaggatata cgcgatgatt aaaagagtcg gaatccgaca
     4081 cctagaatcc tagggcgggt ctggctttag tatatggcga gaggcgagaa aagaggtttc
     4141 aagatcaagg tcgcatgtca tacatgtatt taggaaggga tctttgctgc gcgggtctgc
     4201 gctcaaagta ccggcgtttg gagtttcaac caagtctttc gtttatgcac tgccgcgttg
     4261 ctgtcgatgt tggacctgaa gggctgggga gttgatttgc tcaaaccgtg tatagcagac
     4321 aaagacaacc tggcattcgc gacgatacat gtataggtac gctttacgtc tttcaacccg
     4381 acgtcctcgt tgttctggtc gtgaagtaca tgaatatggt agacgcaacc cctctgcgtc
     4441 tgacgtaacc aagcacataa actgctacgt aaatcgattg acctcgatga agtgatatta
     4501 tggcctgatg tcaatgcagg ctgcaggaca tcaatcagaa ctgcgtatga aggcgacaga
     4561 tggcgacacc ctcggcatga ttttgaccgt cgaatgaagc tcacgtcact ctggcgtggt
     4621 cgagctggaa cagatggatg gacggaatga accttgaagg ttacatatca acggctgctt
     4681 gagctgttca ggtcgggttc caactgttac agaggctcca tatcccccac cacagctgcc
     4741 agcgactgaa accccccctg taacgaaggc aatcgattgc tgtaatctat ccggattgct
     4801 cacgacccca cctcgcccct ccaagcttgc ctatcgttgc agcttgattg tgcgtgcttt
     4861 ttctttccca gagcatctcg gcttgatttt ctctctctct ctttcgtctc ctccatataa
     4921 cttcttcatc acgatttccc taacagcgcg actcttttct cgctatccac gccgatttcc
     4981 tgccaaaatg aagaagccgg agttgactgc aacctccgtt gagaagttcc tgatcgagaa
     5041 gttcgattca gtcagcgatt tgatgcagct ttcggagggt gaagaatcgc gcgcattctc
     5101 atttgacgtt ggcggccgag gctacgtctt gcgagttaac agctgcgccg acggtttcta
     5161 caaggatcgc tatgtctacc gccattttgc ttcagctgcg ctgccaattc cagaagtcct
     5221 ggatatcggc gagtttagcg agtcccttac ctattgcatc tcaagacgag cccaaggtgt
     5281 cacgctccag gacctgcccg agactgagct gccagccgtt ttgcagcccg tcgccgaggc
     5341 gatggacgcc attgctgcgg cagacctcag ccaaacgtcc ggcttcggcc cctttggccc
     5401 tcagggcatc ggccaataca ccacctggcg tgattttatc tgtgccatcg ccgacccaca
     5461 cgtgtaccac tggcagactg tgatggacga caccgtctcc gcgagcgtcg cccaagccct
     5521 ggacgaactg atgctctggg ccgaagattg cccagaggtc cgacacctcg ttcacgctga
     5581 cttcggaagc aacaatgtcc tcaccgataa cggtcgcatc accgccgtta tcgactggtc
     5641 tgaggccatg ttcggagatt cccagtacga ggtcgcaaac atctttttct ggcgaccttg
     5701 gttggcttgt atggagcaac agacccgata tttcgagcga cgccatcctg aactggcggg
     5761 ctctcctcgc ctccgcgcct atatgctgcg aatcggtctg gaccagctgt accagtcact
     5821 cgtcgacgga aactttgacg acgccgcctg ggcccaaggt cgatgtgacg cgatcgtgcg
     5881 cagcggcgct ggaactgttg gacgcacgca gatcgcacgc cgctctgcgg cagtgtggac
     5941 tgatggctgc gtcgaggtcc tcgctgactc aggtaaccgt agaccctcca cgcgcggaga
     6001 tcgcgagatg ggcgaggcca actaaagctc cgtggcgaaa gcctgacgca ccggtagatt
     6061 cttggtgagc ccgtatcatg acggcggcgg gagctacatg gccccgggtg atttattttt
     6121 tttgtatcta cttctgaccc ttttcaaata tacggtcaac tcatctttca ctggagatgc
     6181 ggcctgcttg gtattgcgat gttgtcagct tggcaaattg tggctttcga aaacacaaaa
     6241 cgattcctta gtagccatgc attttaagat aacggaatag aagaaagagg aaattaaaaa
     6301 aaaaaaaaaa acaaacatcc cgttcataac ccgtagaatc gccgctcttc gtgtatccca
     6361 gtaccacggc aaaggtattt catgatcgtt caatgttgat attgttcccg ccagtatggc
     6421 tccaccccca tctccgcgaa tctcctcttc tcgaacgcgg tagtggcgcg ccaattggta
     6481 atgacccata gggagacaaa cagcataata gcaacagtgg aaattagtgg cgcaataatt
     6541 gagaacacag tgagaccata gctggcggcc tggaaagcac tgttggagac caacttgtcc
     6601 gttgcgaggc caacttgcat tgctgtcaag acgatgacaa cgtagccgag gaccgtcaca
     6661 agggacgcaa agttgtcgcg gatgaggtct ccgtagatgg catagccggc aatccgagag
     6721 tagcctctca acaggtggcc ttttcgaaac cggtaaacct tgttcagacg tcctagccgc
     6781 agctcaccgt accagtatcg aggattgacg gcagaatagc agtggctctc caggatttga
     6841 ctggacaaaa tcttccagta ttcccaggtc acagtgtctg gcagaagtcc cttctcgcgt
     6901 gcgagtcgaa agtcgctata gtgcgcaatg agagcacagt aggagaatag gaacccgcga
     6961 gcacattgtt caatctccac atgaattgga tgactgctgg gcagaatgtg ctgcctccaa
     7021 aatcctgcgt ccaacagata ctctggcagg ggcttcagat gaatgcctct gggcccccag
     7081 ataagatgca gctctggatt ctcggttacg atgatatcgc gagagagcac gagttggtga
     7141 tggaggggac gaggaggcat aggtcggccg caggcccata accagtcttg cacagcattg
     7201 atcttcctca cgaggagctc ctgatgcaga aactcctcca tgttgctgat tgggttgaga
     7261 atttcatcgc tcctggatcg tatggttgct ggcaagaccc tgcttaaccg tgccgtgtca
     7321 tggtcatctc tggtggcttc gtcgctggcc tgtctttgca attcgacagc aaatggtgga
     7381 gatctctcta tcgtgacagt catggtagcg atagctaggt gtcgttgcac gcacataggc
     7441 cgaaatgcga agtggaaaga atttcccggc gcggaatgaa gtctcgtcat tttgtactcg
     7501 tactcgacac ctccaccgaa gtgttctcga gatcggatgc cgggaccgac gagtgcagag
     7561 gcgtgcaagc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7621 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7681 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7741 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7801 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7861 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7921 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7981 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8041 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8101 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8161 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8221 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8281 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8341 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8401 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8461 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8521 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8581 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8641 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8701 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8761 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8821 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8881 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8941 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     9001 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9061 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9121 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     9181 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9241 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9301 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9361 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9421 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9481 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9541 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9601 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9661 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9721 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     9781 aaaataggcg tatcacgagg ccctttcgtc
//
