LOCUS       sequence-446698-        5794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for 10x-MBP-GlcR (MGlcR). GlcR is a 
            glycolate-responsive transcriptional repressor and the gene product 
            of pden4400 from Paracoccus denitrificans, codon optimized for E. 
            coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5794)
  AUTHORS   Barthel S, Brenker L, Diehl C, Bohra N, Giaveri S, Paczia N, Erb TJ
  TITLE     In vitro transcription-based biosensing of glycolate for prototyping
            of a complex enzyme cascade.
  JOURNAL   Synth Biol (Oxf). 2024 Sep 20;9(1):ysae013. doi: 
            10.1093/synbio/ysae013. eCollection 2024.
  PUBMED    39399720
REFERENCE   2  (bases 1 to 5794)
  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: "Synth Biol 
            (Oxf)."; date: "2024-09-20"; volume: "9(1):ysae013. doi"; pages: " 
            10.1093/synbio/ysae013. eCollection 2024"
COMMENT     Sequence Label: pTE5418_pMBP-GlcR
FEATURES             Location/Qualifiers
     source          1..5794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..116
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     200..219
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        200..218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    219..243
                     /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             258..280
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             293..322
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             332..1429
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1403..1426
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1478..1489
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1523..1534
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      2389..2446
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2470..2487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2641..3229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2721..2740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3412..3506)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3527..4186)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4101..4120
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4348..4366
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4370..4413)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4416..4437
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     5569..5588
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5608..5685)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
       61 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
      121 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
      181 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
      241 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg gccatcatca
      301 tcatcatcat catcatcatc acagcagcgg caaaatcgaa gaaggtaaac tggtaatctg
      361 gattaacggc gataaaggct ataacggcct cgctgaagtc ggtaagaaat tcgagaaaga
      421 taccggaatt aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt
      481 tgcggcaact ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta
      541 cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta
      601 tccgtttacc tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt
      661 tgaagcgtta tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga
      721 agagatcccg gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa
      781 cctgcaagaa ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa
      841 gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc
      901 gggtctgacc ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta
      961 ctccatcgca gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg
     1021 ggcatggtcc aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt
     1081 caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag
     1141 tccgaacaaa gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct
     1201 ggaagcggtt aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga
     1261 gttggtgaaa gatccgcgta ttgccgccac tatggaaaac gcccagaaag gtgaaatcat
     1321 gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc
     1381 cgccagcggt cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc
     1441 gaacaacaac aacaataaca ataacaacaa cctcgggatc gaaggtcgtc atgggtctcg
     1501 aatgatgggt agctctggcc acatcgaagg tcgtcacatg accaacgcgt ctgtaaaaag
     1561 cgaacgcgcg gctgaagccg tcgcccaaca tctggaaagc ctgatcctgg agggttctct
     1621 gcgcccggac gaacgcctcc tgccggaacg cgaactggcg gaacgtctga acgtttcccg
     1681 cagcaccctg cgtgatggtc tgaaaattct ggaggaacgt ggcctgctca ccagcgcggg
     1741 tggccgtggt tctcgcgttg cagcgctggg caaagctgca attaccgatc cgctcatcgc
     1801 gatgctggtt cgccacgcag aagcggcaga tgactacctg gagttccgtg gcatcgttga
     1861 gtctgcggct gccgcactgg cggctcagcg cgctaccgaa gttgaactgg cgcgtatccg
     1921 tgattgcctg ggccgtatgg accgtgctca cgctcgtgcc gatactggtg aggaagcgga
     1981 agcggacacc gaactgcacc tcctgatcta cgaggctagc cataacatga ccctgctcca
     2041 gattatgcag gccctggcgg gtgttctgcg ttccgacgtg atccagaacc gttcccgtct
     2101 gttcgcggtt ccggcaatcc gcgagctcct gcgtcaacag caccgcgcga tcgcggaagc
     2161 gattctggcc cgtgatgcgc aagctgcgcg tcaggcggct ggcgaacaca ttggctacct
     2221 gcgtcaagca agccgcgaaa tccgcgaagc tgaagctcgt ctggacctga gcctgcgtcg
     2281 cctcaacggt ggcggtgcgg gtacccgtgc cgcaggcacc aaatctactc cggcggcaga
     2341 aggttaagct ttgagaccac tcatcgcgac tagtagcggc cgctgcagtc cggcaaaaaa
     2401 gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat
     2461 gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2521 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2581 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2641 tttccacagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2701 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2761 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2821 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2881 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2941 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3001 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3061 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3121 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3181 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3241 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3301 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3361 atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg gattctcacc
     3421 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     3481 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct
     3541 gccactcgtc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     3601 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     3661 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     3721 aactggtgaa actcacccag ggattggctg atacgaaaaa catattctca ataaaccctt
     3781 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     3841 actgccggaa atcgtcgtgg tattcactcc agagtgatga aaacgtttca gtttgctcat
     3901 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     3961 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     4021 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     4081 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     4141 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     4201 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     4261 gaaagttgga acctcttacg tgcccgatca actcgagtgc cacctgacgt ctaagaaacc
     4321 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa
     4381 aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct tctagagcga
     4441 tgagtctggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     4501 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa actgtcgtgc
     4561 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     4621 ggtggttttt cttttcacca gtgagactgg caacagctga ttgcccttca ccgcctggcc
     4681 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     4741 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     4801 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     4861 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     4921 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     4981 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5041 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5101 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     5161 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     5221 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     5281 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     5341 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     5401 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     5461 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     5521 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     5581 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     5641 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     5701 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     5761 ccgccgccgc aaggaatggt gcatgcaagg agat
//
