LOCUS       sequence-92100-e        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of 71B2 (Q9M156) under T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  TITLE     Anderson Lab Glucosyltransferase plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7083)
  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            "Unpublished"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-72B1
FEATURES             Location/Qualifiers
     source          1..7083
                     /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(1838..1856)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1852..1899
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2074..2079,2080..2096,2097..2102))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2129..2147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2215..2319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2320..2388,2389..3180)
                     /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(2538..2557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3351..3939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3840..3859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4093..4110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4125..4267
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4193..4212)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4353..4375
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4369..4560)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5872..6954)
                     /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     6916..6935
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6955..7032)
                     /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 taccatggag gaatccaaaa
      361 cacctcacgt tgcgatcata ccaagtccgg gaatgggtca tctcatacca ctcgtcgagt
      421 ttgctaaacg actcgtccat cttcacggcc tcaccgttac cttcgtcatc gccggcgaag
      481 gtccaccatc aaaagctcag agaaccgtcc tcgactctct cccttcttca atctcctccg
      541 tctttctccc tcctgttgat ctcaccgatc tctcttcgtc cactcgcatc gaatctcgga
      601 tctccctcac cgtgactcgt tcaaacccgg agctccggaa agtcttcgac tcgttcgtgg
      661 agggaggtcg tttgccaacg gcgctcgtcg tcgatctctt cggtacggac gctttcgacg
      721 tggccgtaga atttcacgtg ccaccgtata ttttctaccc aacaacggcc aacgtcttgt
      781 cgttttttct ccatttgcct aaactagacg aaacggtgtc gtgtgagttc agggaattaa
      841 ccgaaccgct tatgcttcct ggatgtgtac cggttgccgg gaaagatttc cttgacccgg
      901 cccaagaccg gaaagacgat gcatacaaat ggcttctcca taacaccaag aggtacaaag
      961 aagccgaagg tattcttgtg aataccttct ttgagctaga gccaaatgct ataaaggcct
     1021 tgcaagaacc gggtcttgat aaaccaccgg tttatccggt tggaccgttg gttaacattg
     1081 gtaagcaaga ggctaagcaa accgaagagt ctgaatgttt aaagtggttg gataaccagc
     1141 cgctcggttc ggttttatat gtgtcctttg gtagtggcgg taccctcaca tgtgagcagc
     1201 tcaatgagct tgctcttggt cttgcagata gtgagcaacg gtttctttgg gtcatacgaa
     1261 gtcctagtgg gatcgctaat tcgtcgtatt ttgattcaca tagccaaaca gatccattga
     1321 catttttacc accgggattt ttagagcgga ctaaaaaaag aggttttgtg atcccttttt
     1381 gggctccaca agcccaagtc ttggcgcatc catccacggg aggattttta actcattgtg
     1441 gatggaattc gactctagag agtgtagtaa gcggtattcc acttatagca tggccattat
     1501 acgcagaaca gaagatgaat gcggttttgt tgagtgaaga tattcgtgcg gcacttaggc
     1561 cgcgtgccgg ggacgatggg ttagttagaa gagaagaggt ggctagagtg gtaaaaggat
     1621 tgatggaagg tgaagaaggc aaaggagtga ggaacaagat gaaggagttg aaggaagcag
     1681 cttgtagggt gttgaaggat gatgggactt cgacaaaagc acttagtctt gtggccttaa
     1741 agtggaaagc ccacaaaaaa gagttagagc aaaatggcaa ccactaaaga tccggctgct
     1801 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1861 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1921 ggatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
     1981 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
     2041 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc
     2101 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2521 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     2881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3481 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4141 ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct
     4201 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4261 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4321 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4381 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4441 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4501 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4561 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4621 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4681 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     4741 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     4801 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     4861 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     4921 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     4981 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     5041 gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     5101 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
     5161 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc
     5221 catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat
     5281 gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca
     5341 gtgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg
     5401 tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg
     5461 agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag
     5521 cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     5581 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     5641 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     5701 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     5761 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     5821 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5881 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5941 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     6001 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     6061 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     6121 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     6181 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     6241 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     6301 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     6361 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     6421 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     6481 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     6541 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     6601 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     6661 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     6721 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     6781 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6841 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6901 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6961 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     7021 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     7081 cca
//
