LOCUS       sequence-197169-        6996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a beta-glucosidase gene from Thermomicrobium roseum 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 6996)
  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 6996)
  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_BGL1
FEATURES             Location/Qualifiers
     source          1..6996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(109..177,178..969)
                     /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(327..346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1140..1728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1629..1648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1882..1899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1914..2053
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1979..1998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2139..2161
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2155..2346)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3658..4740)
                     /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     4702..4721
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4741..4818)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5024..5043
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5127..5146
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5127..5145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5146..5170
                     /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             5185..5207
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6623..6641)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6637..6684
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6859..6864,6865..6881,6882..6887))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6914..6932)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
       61 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
      361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
      541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
      601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
      661 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
      721 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
      781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
      841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
      901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
      961 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1261 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     1921 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     1981 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2041 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2101 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2161 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2221 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2281 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2341 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2401 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2461 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2521 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2581 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     2641 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     2701 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     2761 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     2821 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     2881 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     2941 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     3001 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     3061 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     3121 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     3181 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     3241 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     3301 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     3361 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     3421 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     3481 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     3541 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     3601 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3721 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     3781 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     3841 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     3901 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     3961 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     4021 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     4081 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     4141 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     4201 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     4261 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     4321 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     4381 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     4441 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     4501 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     4561 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     4621 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     4681 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     4741 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     4801 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     4861 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     4921 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     4981 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     5041 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     5101 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     5161 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgagcg
     5221 tagcgcggtt tccacatggc ttcctgtggg gcactgctac cgccgcatac caaatagagg
     5281 gtgccgtcgg agaagacgga cgggggccgt caatctggga cgccttttcg cacgcccccg
     5341 gcaaaacgta ccaggggcac actggggatg ttgcatgtga ccactatcac cgttggccag
     5401 atgacgtacg tcttttgcgc gaattgggtg cccccgccta tcgcttctca atagcctggc
     5461 ctcgtgtctt ccccgaaggt acaggggctc ctaacgagcc cggccttact ttctatgaac
     5521 ggcttgtaga cgcactgctt gaagccggta taacgccatt tgttacactg tatcattggg
     5581 acctgccgca agcgttacag gaccgggggg gttggtccga acggtccaca gctgaggcgt
     5641 tcgcgagata cgctgaagtg gtaactcgtc ggttaggcga tcgtgtgcgg tactggatca
     5701 cccataacga gccgtgggtg gtggcgtacc tgggtcacta cttaggcgtt catgcaccgg
     5761 gtattcaaga cttacccacc gcgattcgtg ttagtcacga gttacttgta tcacacggac
     5821 tggctgcaca ggcaattcgg gctgcgtcgc cacaggcggt agtcgggatt accctgaatc
     5881 tgagtcctgt cgaaagtgcc tcggatcaac ctgacgaccg cgctgcagca caacttttcg
     5941 atggcttgct gaaccgttgg ttcttagacc ctgtatttgg acggggatac ccacttgatg
     6001 tacgggagcg tttagctcct tatttcgatc ccccagacac cgaccttcca accatagccc
     6061 aaccacttga ctttcttggc attaactatt acgccccggc atttatcaga gctgtacccc
     6121 aagaacaaaa tttgttgggt ttggcgcaat tggagcccaa agcgttagca gacagaggat
     6181 atcagttaac tgacatgggc tggccggttg tccccgaagg tttagaacac ttattgctgc
     6241 gcatcagtag agagtatgcg ccacgcgagt tatttataac cgaaaacggt gccgcctatc
     6301 ctgaccaatt agttgcaggc cgggtacagg acgaccaaag aattgcttat ttagctgcgc
     6361 acttcgctgc cgcttacaga gcagtccggg ccggtgttcc acttaaggga tatttcgtgt
     6421 ggtcccttct ggataacttt gaatgggcgc atggatattc caaacggttc ggcatcgtct
     6481 atgttgatta tgaatcactt tctcgtttcc cgaaagcgag tttcagctgg taccgtgaca
     6541 tcgtcgcaag aaatgctttg gtggaaggtt agggatccgg ctgctaacaa agcccgaaag
     6601 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     6661 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga
     6721 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga
     6781 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta
     6841 actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct
     6901 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     6961 gtttcttaga cgtcaggtgg cacttttcgg ggaaat
//
