LOCUS       sequence-92102-e        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of 89B1 (AT1G73880) under T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  TITLE     Anderson Lab Glucosyltransferase plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7061)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET-89B1
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     155..174
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     258..277
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        258..276
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    277..301
                     /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             316..338
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1816..1834)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1830..1877
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2052..2057,2058..2074,2075..2080))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2107..2125)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2193..2297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2298..2366,2367..3158)
                     /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(2516..2535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3329..3917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3818..3837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4071..4088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4103..4245
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4171..4190)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4331..4353
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4347..4538)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5850..6932)
                     /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     6894..6913
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6933..7010)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
       61 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      121 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      181 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
      241 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
      301 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgaaa gtgaacgagg
      361 aaaacaacaa gccgacaaag acccatgtct taatcttccc atttccggcg caaggtcaca
      421 tgattcccct cctcgacttc acccaccgcc ttgctctccg cggcggcgcc gccttaaaaa
      481 taaccgtcct agtcactcca aaaaaccttc cttttctctc tccgcttctc tccgccgtag
      541 ttaacatcga accacttatc ctcccttttc cctcccaccc ttcaatcccc tccggcgtcg
      601 aaaacgtcca agacttacct ccttcaggct tccctttaat gatccacgcg cttggtaatc
      661 tccacgcgcc gcttatctct tggattactt ctcacccttc tcctccagta gccatcgtat
      721 ctgatttctt ccttggttgg accaaaaacc tcggaatccc tcgtttcgat ttctctccct
      781 ccgctgctat cacttgctgc atactcaata ctctctggat cgaaatgccc accaagatca
      841 acgaagatga cgataacgag atcctccact ttcccaagat cccgaattgt ccaaaatacc
      901 gttttgatca gatctcctct ctttacagaa gttacgttca cggagatcca gcttgggagt
      961 tcataagaga ctcctttaga gataacgtgg cgagttgggg actcgtcgtg aactcgttca
     1021 ccgccatgga aggtgtttat ctcgaacatc ttaagcgaga gatgggccat gatcgtgtat
     1081 gggctgtagg cccaattatt ccgttatctg gggataaccg tggtggcccg acttctgttt
     1141 ctgttgatca cgtgatgtcg tggcttgacg cacgtgagga taaccacgtg gtgtacgtgt
     1201 gctttggaag tcaagtagtt ttgactaaag agcagactct tgcactcgcc tctgggcttg
     1261 agaaaagcgg cgtccatttc atatgggccg taaaggagcc cgttgagaaa gactcaacac
     1321 gtggcaacat cctggacggt ttcgacgatc gcgtggctgg gagaggtctg gtgatcagag
     1381 gatgggctcc acaagtagct gtgctacgtc accgagccgt tggcgcgttt ttaacgcact
     1441 gtggttggaa ctctgtggtg gaggcggttg tcgccggcgt tttgatgctg acgtggccga
     1501 tgagagctga ccagtacact gacgcgtctc tggtggttga tgagttgaaa gtaggtgtgc
     1561 gtgcttgcga aggacctgac acggtgcctg acccggacga gttagctcga gttttcgctg
     1621 attccgtgac cggaaatcaa acggagagga tcaaagccgt ggagctgagg aaagcagcgt
     1681 tggatgcgat tcaagaacgt gggagctcag tgaatgattt agatggattt atccaacatg
     1741 tcgttagttt aggactaaac aaatgagatc cggctgctaa caaagcccga aaggaagctg
     1801 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1861 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatcccgca agaggcccgg
     1921 cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac
     1981 gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga
     2041 taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac
     2101 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2161 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2221 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2281 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2341 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2401 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2461 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2521 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2581 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2641 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2701 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2761 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2821 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2881 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2941 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3001 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3061 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3121 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4021 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4081 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4141 tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4201 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     4261 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4321 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4381 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     4441 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     4501 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     4561 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     4621 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4681 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4741 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4801 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4861 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4921 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4981 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     5041 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     5101 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     5161 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     5221 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     5281 ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt aggctggtaa
     5341 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     5401 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     5461 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     5521 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     5581 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     5641 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     5701 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     5761 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5821 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5881 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5941 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     6001 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     6061 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     6121 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     6181 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     6241 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     6301 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     6361 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     6421 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     6481 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     6541 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6601 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6661 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6721 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6781 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6841 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6901 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6961 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     7021 acgctctccc ttatgcgact cctgcattag gaagcagccc a
//
