LOCUS       sequence-354318-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier B8MMC0.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  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 6851)
  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: B8MMC0
FEATURES             Location/Qualifiers
     source          1..6851
                     /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             1571..1600
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1653..1671)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1667..1714
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1751..2206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1838..1857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2048..2069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2232..2336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2337..2405,2406..3197)
                     /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(2555..2574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3368..3956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3857..3876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4110..4127
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4142..4281
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4207..4226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4367..4389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4383..4574)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5383..6465)
                     /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     6427..6446
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6466..6543)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6749..6768
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gagccagcaa aataaaggcg ttcagatctc
      121 gggtgcagag gtagcacagc ataacaataa ggactcttgc tggattgcag ttcgtggcaa
      181 agtgtatgat gtaactgatt ttcttgagga acatccaggc ggggctcgtt tgttgcttaa
      241 atgtgcgggc cgcgacgcca ccgaggacta tgacgcgatc cacccggctc agctcattga
      301 ggaaagcctg ccggattcgg ccttcaaagg caccgtggat cctagcgagt tggatcgtcc
      361 aaagctggcg gacaaaagtg tgcgtaacgc ccagaaagaa ggtaagaagg atccagattc
      421 gccgccgcct ttaagtgcga tgttgaatgt acgcgatttt gaaaaggtcg cggagcgcca
      481 cctggctgaa aacagctggg cctactatac cgcgggcgcg gacgatgagt attcaaaggc
      541 cgaggcggaa ctggcttatc gtaaggtgtt attccgcccg cgtattctgc gcaatgtagg
      601 ccgcgtagat actcgtacgc aaattcttgg ccaagacgta tccctgccga tttatatctc
      661 tgcggtgggc atcgcaaagt ttgcccatcc gcaaggcgaa tgcacgctgg ccgccgcagc
      721 gggacgtgaa ggcattgccc agctggtagc aacgcgttcc tctatgagca tcgaaagcat
      781 tatgaaagcc cgtacgggtg gtccgcaaca gcctatcttc tttcaactct atatgcacaa
      841 agacgccaag atttctgatg ccacgatcct gaaggctgtt aaagcaggcg tgaaaggcat
      901 ttggttaaca gttgacagtc cggtaactgg caagcgcgag cgcgatgaac gtctcaaagc
      961 gaatgtggat gttggggaac aaaatgataa gattggtggt aaaggtaaac cggtacaagg
     1021 ggtggcaaag acactgagca gcactgttag tccgtatctg gattgggaca ccatttctta
     1081 tattcgcaaa ctgactaacc tgccactggt aattaaaggg atccaaagtg tggaagatgc
     1141 cattctggcc cataagcaca aagtcaatgg aattgtgatt tctaatcacg gcggtcgttc
     1201 gcaagatacc gcacaggccc cgctgttgac gttacttgaa attaacaagt acgcgccgca
     1261 aattatcacc gataagaaga tgcaaatctt tattgacggc ggggtgcgcc gtggtaccga
     1321 cgtggtgaaa gccctggcct taggtgctac cgcggtaggt atgggccgcc cgttccttta
     1381 ttctatgagc tcggggtacg gagaggcagg cactcgccgc atgattgaga tcatgcgcga
     1441 ggaaattgag caaaacatgg cgctcgtggg cgctacgaag atcagcgagc tgcgccgcga
     1501 acttctgaac accagccgcc ttgagcgcga tctcacaacg tttattgcaa aactcgcggc
     1561 cgcactcgag catcatcacc atcaccacca ccaccaccac tgagatccgg ctgctaacaa
     1621 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1681 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     1741 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1801 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1861 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1921 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1981 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2041 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2101 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2161 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2281 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2341 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2401 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2461 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2521 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2581 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2641 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2701 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2761 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2821 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2881 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2941 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3001 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3061 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3121 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3181 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3241 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3301 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3481 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4021 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4081 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4141 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     4201 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     4261 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4321 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4381 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     4441 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     4501 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     4561 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     4621 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     4681 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     4741 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     4801 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     4861 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     4921 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     4981 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     5041 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     5101 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     5161 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     5221 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     5281 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     5341 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     5401 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5461 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     5521 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     5581 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     5641 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     5701 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     5761 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     5821 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     5881 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     5941 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     6001 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6061 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6121 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     6181 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     6241 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     6301 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     6361 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     6421 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     6481 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     6541 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     6601 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     6661 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     6721 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     6781 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     6841 tcccgcgaaa t
//
