LOCUS       sequence-192844-        7401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a beta-glucosidase gene from Thermobaculum terrenum in E. 
            coli under the control of the pET system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  AUTHORS   Husser MC, Vo PQN, Sinha H, Ahmadi F, Shih SCC
  TITLE     An Automated Induction Microfluidics System for Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2018 Mar 16;7(3):933-944. doi: 
            10.1021/acssynbio.8b00025. Epub 2018 Mar 8.
  PUBMED    29516725
REFERENCE   2  (bases 1 to 7401)
  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"; doi: 
            "10.1021/acssynbio.8b00025"; journalName: "ACS Synth Biol"; date: 
            "2018-03-16- 16"; volume: "7"; issue: "3"; pages: "933-944"
COMMENT     Sequence Label: pET_BGL2
FEATURES             Location/Qualifiers
     source          1..7401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    38..62
                     /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             77..99
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1917..1935)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1931..1978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2153..2158,2159..2175,2176..2181))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2208..2226)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2294..2398
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2399..2467,2468..3259)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2617..2636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3430..4018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3919..3938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4172..4189
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4204..4346
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4272..4291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4432..4454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4448..4639)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5951..7033)
                     /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     6995..7014
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7034..7111)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7317..7336
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
       61 cccctctaga aataattttg tttaacttta agaaggagat ataccatgga cccgtatgaa
      121 gatccgcgtc tgccggttga agcacgtatt gaagatctgc tgggtcgtat gaccctggaa
      181 gaaaaagcag gtatgctgtt tcataccggt ctgggtatga atccggatgg caccctgcaa
      241 gaaggtgacg gcacctttgg tcgtgcaagc accaccgaac tggttacccg taaactgctg
      301 aatcatttta atgtttgggc agttgcagat cctcgtccga tggcagaatg gtataatcgt
      361 ctgcaggcac tggcagaagg cacccgtctg ggcattccga ttaccattag cagcgatcca
      421 cgtcatagct atagcaataa tccggcagca agcctgtttg caggtcgttt tagcaaatgg
      481 cctgaaccga ttggtctggc agcaattggt gatgaagaac tggttcgtgc atttggtgat
      541 attgcacgtc aagaatatct ggcagttggt attcgtgttg cactgcatcc gatggccgat
      601 ctggcaaccg aaccgcgttg ggcacgcatt gcaggtactt ttggtgaaga tgcacatctg
      661 gcagcccatc tggttgcagc atatatcaaa ggttttcagg gtgaaaatct gggtccgcat
      721 agcgttgcat gtatgaccaa acattttcct ggtggtggtc cgcagaaaga tggcgaagat
      781 cctcattttc cgtatggtcg tgaacaggtt tatccgggta ataactttga gtatcatctg
      841 attccgtttg aagcagcatt tgaagccggt acagcacaga ttatgccgta ttatggtatg
      901 ccggttggcc tgccttttga agaagttggt tttggtttta atcgtgatgt tattgcaggt
      961 ctgctgcgtc agcgttatgg ctttcagggc gttgtttgta ccgattgggg tctgctgacc
     1021 gatcatcgta tgggtgatcg tgttctgagc gcacgtgcat ggggtgttga acatctgagc
     1081 ctggaagatc gtgtgctgaa agcactggat gccggtgttg atcagtttgg tggtgaaagc
     1141 tgtccggaag ttgttgttca gctggttcgt agcggtcgcc tgccggaaga acgtctggat
     1201 gttagcgttc gtcgcctgct gcgtgataaa tttcgtctgg gtctgtttga taacccgttt
     1261 gttgatccgg aagaagcaga acgcgttgtt ggtcaagaag catttgttcg cgcaggcgaa
     1321 gcagcacagc gtcgtagcat tgttctgctg gtgaatggtg aaaccagcag cggtcgtacc
     1381 ctgccgatgc gtgaaggtat tcgtctgtat gttgaaggcg ttgatccgca ggttgcaagc
     1441 cgttatgcac aggttgttga tacaccggaa ggtgcagatg cagcatttat tcgtttacag
     1501 gcaccgtatg aacatcgtga taatctgccg ctggaagcat tttttcatgc aggtaatctg
     1561 agctttccgg aaccggaaca gcgtcgcatt ctggatttac tgcgtcgtgt tccgaccgtt
     1621 gttcagattt atctggatcg tccggcagtt attccggaaa ttgcacgtga aagcgcagca
     1681 ctgctggcag attttggtgc aagtgatgaa gcagttctgg atgtggcatt tggtcgtcat
     1741 aaacctggtg gtcgtctgcc atttgaaatg ccgagcagca tggatgcagt tcgtaaacag
     1801 ctgcctgatg ttccgtgtga tagtgaagat ccactgtttc cgatcggtca tggtctgacc
     1861 ctgtagggat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1921 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1981 gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     2041 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     2101 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     2161 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
     2221 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2281 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2341 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2401 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2461 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2521 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2581 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2641 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2701 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2761 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2821 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2881 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2941 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3001 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3061 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3121 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3181 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3241 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3301 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3361 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3421 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3481 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3541 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3601 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3661 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3721 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3781 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3841 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3901 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3961 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4021 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4081 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4141 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4201 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     4261 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     4321 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4381 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4441 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4501 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4561 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4621 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4681 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4741 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4801 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     4861 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4921 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4981 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     5041 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     5101 cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg
     5161 acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt
     5221 gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca
     5281 ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag
     5341 ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc
     5401 cgtgacgatc agcggtccag tgatcgaagt taggctggta agagccgcga gcgatccttg
     5461 aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat
     5521 cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc
     5581 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     5641 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     5701 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     5761 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     5821 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     5881 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     5941 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6001 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     6061 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     6121 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     6181 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     6241 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     6301 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     6361 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     6421 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     6481 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     6541 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     6601 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     6661 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     6721 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     6781 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     6841 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     6901 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     6961 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     7021 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     7081 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     7141 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     7201 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     7261 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     7321 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     7381 cgtccggcgt agaggatcga g
//
