LOCUS       sequence-354280-        7058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier M7MTF7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7058)
  AUTHORS   Rembeza E, Engqvist MKM
  TITLE     Experimental and computational investigation of enzyme functional 
            annotations uncovers misannotation in the EC 1.1.3.15 enzyme class.
  JOURNAL   PLoS Comput Biol. 2021 Sep 23;17(9):e1009446. doi: 
            10.1371/journal.pcbi.1009446. eCollection 2021 Sep.
  PUBMED    34555022
REFERENCE   2  (bases 1 to 7058)
  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: "PLoS Comput
            Biol."; date: "2021-09-23"; volume: "17(9):e1009446. doi"; pages: " 
            10.1371/journal.pcbi.1009446. eCollection 2021 Sep"
COMMENT     Sequence Label: M7MTF7
FEATURES             Location/Qualifiers
     source          1..7058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1778..1807
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1860..1878)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1874..1921
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1958..2413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2045..2064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2255..2276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2439..2543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2544..2612,2613..3404)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2762..2781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3575..4163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4064..4083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4317..4334
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4349..4488
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4414..4433)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4574..4596
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4590..4781)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5590..6672)
                     /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     6634..6653
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6673..6750)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6956..6975
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggttacccag aataccgtcg aaatcccacg
      121 ctctgtttgg tacggttggg gagaccctgc ccacgccacc tctctttccg caaatgcctt
      181 aacatatttg agcgcaactc tgaagcttgc aagcgttaaa ccagtgcacc caccggctga
      241 cattacagaa gtgcgcattg gggaacccgt gttaaatgcc gaagccctcg ccgaattacg
      301 cgcagcgtgt ggtacagaat atgtctcgac tgagcgtctc gaccgcattg agcatagcgg
      361 cggtaaaagt accgaagatc tgttacgcat tcgcgcgggc gatgcgctgg gagcacctga
      421 tgcagtcatc tttcctgcta gtagtgaaga aattcgtgaa gtactggcag tttgtgcggc
      481 acgtggaatc gcggtaaccg ctttcggcgg cggaacttcg gttgtaggtg gagtatcacc
      541 tcttcgtggc acctttgatg ctgcgattac gctggatatg cgccgtatga ataaaatgct
      601 tcatcttgac ccgcgttcac gcacagcgac atttgaggca ggcttacgtg gtccgggcat
      661 cgaagcagca ctggaacccc atggattcac tttaggacat ttccctcagt ctcaccaaga
      721 ggcgacatta ggtggttacg tggccacgcg ttctgccggt caagcctcaa ccggttacgg
      781 ccgtagcgat gatctcgtaa aatccgtgca tcttgagact ccggctggtc ctctggtggc
      841 gggttcgaca gcaccgggta ccgcagcggg tccgaagctt ttggatgtca tcgtgggtag
      901 cgaaggagca tttggagtga tcacctcggc aaccctgaaa attgttcctg ttcctaccca
      961 taagtcatat ggcagttggt cctttgcctc ttttacggcc ggggctgagg cgattcgccg
     1021 tctgcgtcat gaaggcctca gtggtgatct gccgcatgtt tgtcgtttaa gcgattccga
     1081 tgaaaccgct tcgaccttca aactcggtgg accaaagacg gcggccttgg ggaagtacct
     1141 gagcgtgcgt ggtcaggcca caccagctct ggccctgttc ctttgggaag gtgcgactaa
     1201 agagaccgcc gcccgtaaac gtcgtagcgc ccgtattctg cgtgctgcgg gtggcatccc
     1261 aacaggtcct attcccgccc gtgcctggga acatggccgt ttcagtggtc cgtacttgcg
     1321 tgatgaactt ctgacccgcg gcgtgtatgt agaaacttta gagacctcgg caacatggac
     1381 ctcgttggac agcacctacc acgcagtccg cgaggcaatt ttaaaatccc ttgcggaaca
     1441 tggcggcaca ggctatgttc aaagtcacat ctcacatatt tatccggatg gggcaagctt
     1501 atatttcaca tatcttgcag gcgtggaatc agatggtctg gctcaacacg ctcgtgtaaa
     1561 gcaagctgcg tctaaagcga ttgttgccgc gggctcaacg attacgcatc atcactccgt
     1621 ggggaccgat cacgcgccgt acttaggtgc cgagattggc gcgctgggtc tggacgtact
     1681 gcgcggtatt aagaacactc tggatccggc tggaattatg aatccgggta aactgattcc
     1741 agcagcgatt aaagagcaac cggcggccgc actcgagcat catcaccatc accaccacca
     1801 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1861 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1921 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     1981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2341 tattgattaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2401 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2461 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2521 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2581 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2641 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2701 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2761 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2821 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2881 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2941 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3001 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3061 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     3121 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3181 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3241 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3301 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3361 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3421 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3481 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3541 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3601 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3661 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3721 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3781 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3841 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3901 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3961 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4021 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4081 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4141 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4201 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4261 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4321 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4381 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4441 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4501 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4561 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4621 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     4681 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     4741 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     4801 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     4861 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     4921 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     4981 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     5041 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     5101 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     5161 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     5221 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
     5281 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     5341 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     5401 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     5461 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     5521 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     5581 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5641 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     5701 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     5761 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     5821 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     5881 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     5941 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     6001 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     6061 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     6121 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     6181 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     6241 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     6301 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     6361 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     6421 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     6481 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     6541 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     6601 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     6661 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     6721 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     6781 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     6841 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     6901 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6961 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     7021 gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
