LOCUS       sequence-198935-        8157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a beta-glucosidase gene from Rhodothermus marinus 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 8157)
  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 8157)
  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_BGL3
FEATURES             Location/Qualifiers
     source          1..8157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(121..189,190..981)
                     /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(339..358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1152..1740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1641..1660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1894..1911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1926..2065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1991..2010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2151..2173
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2167..2358)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3670..4752)
                     /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     4714..4733
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4753..4830)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5036..5055
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5139..5158
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5139..5157
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5158..5182
                     /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             5197..5219
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5550..5561
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7796..7814)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7810..7857
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(8032..8037,8038..8054,8055..8060))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8087..8105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
       61 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      121 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      181 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      241 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      301 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      361 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      421 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      481 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      541 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      601 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      661 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      721 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      781 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      841 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      901 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      961 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1021 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1081 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1141 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1201 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1261 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1321 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1381 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1441 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1501 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1561 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1621 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1681 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1741 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1801 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1861 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1921 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     1981 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     2041 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     2101 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2161 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     2221 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     2281 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     2341 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     2401 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     2461 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     2521 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     2581 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     2641 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     2701 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     2761 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     2821 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     2881 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     2941 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     3001 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     3061 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     3121 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     3181 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     3241 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     3301 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     3361 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     3421 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     3481 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     3541 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     3601 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     3661 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3721 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     3781 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     3841 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     3901 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     3961 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4021 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     4081 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     4141 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     4201 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     4261 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     4321 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     4381 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     4441 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     4501 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     4561 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     4621 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     4681 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     4741 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     4801 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     4861 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     4921 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     4981 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5041 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     5101 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
     5161 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
     5221 ataccatgaa acatctggtt accacactgc tgtttggtct ggtttttgca gcagcagttc
     5281 gtgcacagag cagccgtgtt gatagcctgc tggcagcaat gaccctggca gaaaaaatca
     5341 gcctgctgca tggtgcaatt gatccgaccg gtgaagccgg tgcaggttat gttccgggtg
     5401 ttccgcgtct gggtattccg gcaatgcgtc tgaccgatgg tccggcaggt attcgtaccc
     5461 gtcatccggc aaccgcactg ccagcaccgg ttgcactggc agccagcttt gatccggaac
     5521 tggcatatcg ttatggtcat gttatgggta ttgaaggtcg tgcacgtcgt catgaagttc
     5581 tgctgagccc gatggttaat attgttcgtg ttccggaagc aggtcgtaac tttgaaacct
     5641 ttagcgaaga tccgctgctg agcgcagaaa tggttgcagc cgaagtgcgt ggtattcagg
     5701 atgcaggtat gatggcaacc gttaaacatt atgtggccaa caactttgag aatgatcgta
     5761 tgcgtgttaa tgtggttgtt gatgaacgta ccctgcgcga aatttatctg cctggttttg
     5821 aagcagccat taaagcgggt gcagcagccg ttatgtgtgc atataatcgt gtgaatggtc
     5881 cgtatgcctg tgataatgaa atgctgctga ccgatattct gcgtgatgaa tggggctttg
     5941 aaggttgggt tatgaccgat tggtttgcag gtcatagcct ggaaagcctg gttcgtggtc
     6001 tggatcaaga aatgcctggt tataccattc cgtttagcag tccggatatg ccgctggcac
     6061 cggcagtttt tgccgatagc ctgttagccg cagttgaaag cggtcgtatt gatgaagcct
     6121 atgttgatcg tgcagttcgt cgtattctgg ttcagatgga acagtttggc ctgctggatg
     6181 gcgaaagcac ccctccggaa attaacattg aagcacatgc agccgttgca cgtgaagttg
     6241 cagaagcagg cgctgttctg ctgcgtaatg aaaatcagac cctgccgctg agcgaacgtg
     6301 atctgcagca tctggttgtt attggtccga ccgcaacacg tccgctgatt ggtggtggcg
     6361 gtagcagtcg tgttcagccg tttcgtacca caagcactct ggcagcactg caagaactgg
     6421 caggtccgca ggcacagatt cgttatgtgc caggtattga tctggatggt atgccggttc
     6481 cgagcagcgc actgcgtaca ccggatggtc agcctggtct gctgcgccag ggtgcagatg
     6541 gtacaaccca ggttgatgca cagctggatt ttaccggtga acgtgcactg cctccgggta
     6601 gccagtggac ctggaccggt acactgaccg caccgaccgc aggcgtttat gaactgaaac
     6661 tgcagacagc cggtggtgtt ggcaccctga gcattgatgg ccagccggtt ctgcgtaccg
     6721 gtatgttttt tagtgatgca agcctgattc cgacagcaga tagtctggaa aatgcaacct
     6781 atcgtattga actgcaggca ggccagcagc tgagcctgac cgttcagatt agcggtcagg
     6841 tgccgagcct gccgtttctg cctgccggtc agaatccggt tcaggttcgt ctggcatggg
     6901 ttacaccgga acgtcgtgca gcctttctgg aagaagcagc ggaagcagca cgtgcagcac
     6961 atgccgcaat tgtttttgtt tatgaagagg gcaccgaagg tcgcgatcgt gaaaccctgg
     7021 cactgcgtcc ggatcaggat gccctggttg aagcagttgc cgcagcaaat ccgcgtacca
     7081 ccgttgttat gaatgttggt gcaccgacac tgatgccgtg ggcagaacgt gttggtgcca
     7141 ttctgctgat gtggtatccg ggtcaagaag gtggttgggc caccgcagat gtgctgctgg
     7201 gtcgtgcaaa tccggcaggc cgtctgccgg ttacctttcc gcgtcgtgcc gaagatgctc
     7261 cgaccgccag tccggaacgt tatcctggtg ttgatctgac agcacgttat gatgaaggca
     7321 tttttgttgg ttatcgttgg tatgatgccc agcagattga accgctgttt ccgtttggtc
     7381 atggtctgag ctataccacc tttgcctatg aaaatctgcg tgttgaacct gatggtgatg
     7441 gttttgttgt tcgttttgtg gtgcgtaata ccggtgatcg cgcaggtagt gatgttccgc
     7501 aggtttatct gggtccgcct gaaaatccgc ctgttccgat ggccgttcgt cagctggttg
     7561 gttttcgtcg tgttacactg gcacctggtg aagcacaaga agttaccgtt cgtatcgatg
     7621 gtcgtgccct gagctattgg agcgttgaag atcatgcctg ggttaaagca accggtcgtc
     7681 gtaccctgta tgtgggtgca agcgcacgcg atctgcgcct gcagaccgaa attgatgttc
     7741 agtagggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     7801 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     7861 aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     7921 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     7981 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     8041 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
     8101 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcact
//
