LOCUS       sequence-197171-        7398 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 7398)
  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 7398)
  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..7398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    22..46
                     /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             61..83
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1901..1919)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1915..1962
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2137..2142,2143..2159,2160..2165))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2192..2210)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2278..2382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2383..2451,2452..3243)
                     /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(2601..2620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3414..4002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3903..3922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4156..4173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4188..4327
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4253..4272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4413..4435
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4429..4620)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5932..7014)
                     /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     6976..6995
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7015..7092)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7298..7317
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
       61 tttgtttaac tttaagaagg agatatacca tggacccgta tgaagatccg cgtctgccgg
      121 ttgaagcacg tattgaagat ctgctgggtc gtatgaccct ggaagaaaaa gcaggtatgc
      181 tgtttcatac cggtctgggt atgaatccgg atggcaccct gcaagaaggt gacggcacct
      241 ttggtcgtgc aagcaccacc gaactggtta cccgtaaact gctgaatcat tttaatgttt
      301 gggcagttgc agatcctcgt ccgatggcag aatggtataa tcgtctgcag gcactggcag
      361 aaggcacccg tctgggcatt ccgattacca ttagcagcga tccacgtcat agctatagca
      421 ataatccggc agcaagcctg tttgcaggtc gttttagcaa atggcctgaa ccgattggtc
      481 tggcagcaat tggtgatgaa gaactggttc gtgcatttgg tgatattgca cgtcaagaat
      541 atctggcagt tggtattcgt gttgcactgc atccgatggc cgatctggca accgaaccgc
      601 gttgggcacg cattgcaggt acttttggtg aagatgcaca tctggcagcc catctggttg
      661 cagcatatat caaaggtttt cagggtgaaa atctgggtcc gcatagcgtt gcatgtatga
      721 ccaaacattt tcctggtggt ggtccgcaga aagatggcga agatcctcat tttccgtatg
      781 gtcgtgaaca ggtttatccg ggtaataact ttgagtatca tctgattccg tttgaagcag
      841 catttgaagc cggtacagca cagattatgc cgtattatgg tatgccggtt ggcctgcctt
      901 ttgaagaagt tggttttggt tttaatcgtg atgttattgc aggtctgctg cgtcagcgtt
      961 atggctttca gggcgttgtt tgtaccgatt ggggtctgct gaccgatcat cgtatgggtg
     1021 atcgtgttct gagcgcacgt gcatggggtg ttgaacatct gagcctggaa gatcgtgtgc
     1081 tgaaagcact ggatgccggt gttgatcagt ttggtggtga aagctgtccg gaagttgttg
     1141 ttcagctggt tcgtagcggt cgcctgccgg aagaacgtct ggatgttagc gttcgtcgcc
     1201 tgctgcgtga taaatttcgt ctgggtctgt ttgataaccc gtttgttgat ccggaagaag
     1261 cagaacgcgt tgttggtcaa gaagcatttg ttcgcgcagg cgaagcagca cagcgtcgta
     1321 gcattgttct gctggtgaat ggtgaaacca gcagcggtcg taccctgccg atgcgtgaag
     1381 gtattcgtct gtatgttgaa ggcgttgatc cgcaggttgc aagccgttat gcacaggttg
     1441 ttgatacacc ggaaggtgca gatgcagcat ttattcgttt acaggcaccg tatgaacatc
     1501 gtgataatct gccgctggaa gcattttttc atgcaggtaa tctgagcttt ccggaaccgg
     1561 aacagcgtcg cattctggat ttactgcgtc gtgttccgac cgttgttcag atttatctgg
     1621 atcgtccggc agttattccg gaaattgcac gtgaaagcgc agcactgctg gcagattttg
     1681 gtgcaagtga tgaagcagtt ctggatgtgg catttggtcg tcataaacct ggtggtcgtc
     1741 tgccatttga aatgccgagc agcatggatg cagttcgtaa acagctgcct gatgttccgt
     1801 gtgatagtga agatccactg tttccgatcg gtcatggtct gaccctgtag ggatccggct
     1861 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     1921 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     1981 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     2041 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     2101 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     2161 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2221 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2281 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2341 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2401 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2461 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2521 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2581 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2641 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2701 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2761 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2821 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2881 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     2941 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3001 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3061 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3121 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3181 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3241 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3301 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3361 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3421 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3481 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3541 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3601 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3661 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3721 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3781 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3841 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3901 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3961 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4021 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4081 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4141 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4201 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     4261 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4321 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4381 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4441 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4501 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4561 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4621 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4681 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4741 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     4801 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     4861 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     4921 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     4981 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     5041 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     5101 gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     5161 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
     5221 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc
     5281 catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat
     5341 gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca
     5401 atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg
     5461 tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg
     5521 agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag
     5581 cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     5641 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     5701 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     5761 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     5821 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     5881 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5941 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6001 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     6061 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     6121 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     6181 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     6241 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     6301 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     6361 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     6421 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     6481 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     6541 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     6601 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     6661 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     6721 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     6781 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     6841 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6901 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6961 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     7021 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     7081 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     7141 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     7201 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     7261 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     7321 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     7381 agatctcgat cccgcgaa
//
