LOCUS       sequence-192843-        8160 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 8160)
  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 8160)
  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..8160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(71..139,140..931)
                     /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(289..308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1102..1690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1591..1610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1844..1861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1876..2018
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1944..1963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2104..2126
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2120..2311)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3623..4705)
                     /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     4667..4686
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4706..4783)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4989..5008
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5092..5111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5092..5110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5111..5135
                     /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             5150..5172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5503..5514
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7749..7767)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7763..7810
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7985..7990,7991..8007,8008..8013))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8040..8058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8126..8160,1..70)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
       61 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      121 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      181 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      241 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      301 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      361 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      421 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      481 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      541 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      601 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      661 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      721 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      781 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      841 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      901 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
      961 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1021 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1081 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1141 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1201 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     1261 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1321 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1381 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1441 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1501 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1561 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1621 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1681 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1741 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1801 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1861 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1921 ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     1981 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2041 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2101 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2161 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     2221 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     2281 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     2341 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2401 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     2461 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     2521 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     2581 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     2641 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     2701 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     2761 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc
     2821 tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc
     2881 tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc
     2941 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
     3001 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
     3061 ggcataaatc gccgtgacga tcagcggtcc agtgatcgaa gttaggctgg taagagccgc
     3121 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     3181 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     3241 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
     3301 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     3361 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     3421 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     3481 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     3541 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3601 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3661 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3721 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3781 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3841 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3901 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3961 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4021 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4081 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4141 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     4201 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     4261 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4321 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     4381 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     4441 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4501 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4561 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4621 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4681 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4741 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     4801 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     4861 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     4921 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     4981 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     5041 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
     5101 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     5161 ttaagaagga gatataccat gaaacatctg gttaccacac tgctgtttgg tctggttttt
     5221 gcagcagcag ttcgtgcaca gagcagccgt gttgatagcc tgctggcagc aatgaccctg
     5281 gcagaaaaaa tcagcctgct gcatggtgca attgatccga ccggtgaagc cggtgcaggt
     5341 tatgttccgg gtgttccgcg tctgggtatt ccggcaatgc gtctgaccga tggtccggca
     5401 ggtattcgta cccgtcatcc ggcaaccgca ctgccagcac cggttgcact ggcagccagc
     5461 tttgatccgg aactggcata tcgttatggt catgttatgg gtattgaagg tcgtgcacgt
     5521 cgtcatgaag ttctgctgag cccgatggtt aatattgttc gtgttccgga agcaggtcgt
     5581 aactttgaaa cctttagcga agatccgctg ctgagcgcag aaatggttgc agccgaagtg
     5641 cgtggtattc aggatgcagg tatgatggca accgttaaac attatgtggc caacaacttt
     5701 gagaatgatc gtatgcgtgt taatgtggtt gttgatgaac gtaccctgcg cgaaatttat
     5761 ctgcctggtt ttgaagcagc cattaaagcg ggtgcagcag ccgttatgtg tgcatataat
     5821 cgtgtgaatg gtccgtatgc ctgtgataat gaaatgctgc tgaccgatat tctgcgtgat
     5881 gaatggggct ttgaaggttg ggttatgacc gattggtttg caggtcatag cctggaaagc
     5941 ctggttcgtg gtctggatca agaaatgcct ggttatacca ttccgtttag cagtccggat
     6001 atgccgctgg caccggcagt ttttgccgat agcctgttag ccgcagttga aagcggtcgt
     6061 attgatgaag cctatgttga tcgtgcagtt cgtcgtattc tggttcagat ggaacagttt
     6121 ggcctgctgg atggcgaaag cacccctccg gaaattaaca ttgaagcaca tgcagccgtt
     6181 gcacgtgaag ttgcagaagc aggcgctgtt ctgctgcgta atgaaaatca gaccctgccg
     6241 ctgagcgaac gtgatctgca gcatctggtt gttattggtc cgaccgcaac acgtccgctg
     6301 attggtggtg gcggtagcag tcgtgttcag ccgtttcgta ccacaagcac tctggcagca
     6361 ctgcaagaac tggcaggtcc gcaggcacag attcgttatg tgccaggtat tgatctggat
     6421 ggtatgccgg ttccgagcag cgcactgcgt acaccggatg gtcagcctgg tctgctgcgc
     6481 cagggtgcag atggtacaac ccaggttgat gcacagctgg attttaccgg tgaacgtgca
     6541 ctgcctccgg gtagccagtg gacctggacc ggtacactga ccgcaccgac cgcaggcgtt
     6601 tatgaactga aactgcagac agccggtggt gttggcaccc tgagcattga tggccagccg
     6661 gttctgcgta ccggtatgtt ttttagtgat gcaagcctga ttccgacagc agatagtctg
     6721 gaaaatgcaa cctatcgtat tgaactgcag gcaggccagc agctgagcct gaccgttcag
     6781 attagcggtc aggtgccgag cctgccgttt ctgcctgccg gtcagaatcc ggttcaggtt
     6841 cgtctggcat gggttacacc ggaacgtcgt gcagcctttc tggaagaagc agcggaagca
     6901 gcacgtgcag cacatgccgc aattgttttt gtttatgaag agggcaccga aggtcgcgat
     6961 cgtgaaaccc tggcactgcg tccggatcag gatgccctgg ttgaagcagt tgccgcagca
     7021 aatccgcgta ccaccgttgt tatgaatgtt ggtgcaccga cactgatgcc gtgggcagaa
     7081 cgtgttggtg ccattctgct gatgtggtat ccgggtcaag aaggtggttg ggccaccgca
     7141 gatgtgctgc tgggtcgtgc aaatccggca ggccgtctgc cggttacctt tccgcgtcgt
     7201 gccgaagatg ctccgaccgc cagtccggaa cgttatcctg gtgttgatct gacagcacgt
     7261 tatgatgaag gcatttttgt tggttatcgt tggtatgatg cccagcagat tgaaccgctg
     7321 tttccgtttg gtcatggtct gagctatacc acctttgcct atgaaaatct gcgtgttgaa
     7381 cctgatggtg atggttttgt tgttcgtttt gtggtgcgta ataccggtga tcgcgcaggt
     7441 agtgatgttc cgcaggttta tctgggtccg cctgaaaatc cgcctgttcc gatggccgtt
     7501 cgtcagctgg ttggttttcg tcgtgttaca ctggcacctg gtgaagcaca agaagttacc
     7561 gttcgtatcg atggtcgtgc cctgagctat tggagcgttg aagatcatgc ctgggttaaa
     7621 gcaaccggtc gtcgtaccct gtatgtgggt gcaagcgcac gcgatctgcg cctgcagacc
     7681 gaaattgatg ttcagtaggg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     7741 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     7801 gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc
     7861 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     7921 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     7981 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
     8041 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     8101 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
//
