LOCUS       sequence-316425-        6420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier F6CI10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  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 6420)
  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: F6CI10
FEATURES             Location/Qualifiers
     source          1..6420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1048..1077
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1130..1148)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1144..1191
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1228..1683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1315..1334)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1525..1546
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1710..1814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1815..1883,1884..2675)
                     /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(2033..2052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2846..3434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3335..3354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3588..3605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3620..3762
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3688..3707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3848..3870
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3864..4055)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4864..5946)
                     /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     5908..5927
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5947..6024)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6230..6249
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6333..6352
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6333..6351
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6352..6376
                     /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             6391..6413
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgaactgga aagagttacg cgaggccgca cgcgaaaagt tgaagggcta ttgccgtgtc
       61 tgcccctcat gtgacgggcg cgcgtgtgct ggagaagttc caggaatggg tggaacagga
      121 acgggcgcgt cttttaaggc taacatcgag gcattggcac gctatcgctt aaacttacgt
      181 acattacacg gcgcaaagaa cccagacacc agcttaaccc ttttcgggaa agaactctct
      241 actccaatct tagctgcacc catgaccggt gtcacgtata atatgggcgg agctcttact
      301 gagcgtgaat ttattggcat gatcatggca ggtgcacgcc aagcgggtac gctcggtatg
      361 tctggagatg gcggcgatcc ggaatattat aattcgggcc tcgaagcaat cgctgctgaa
      421 ggtggccacg gtattcccgt gattaagcca cgcgaacaac aagccgtgat cgaacgcatc
      481 cgccgcgccg aagaagccgg agccccagcg gtcggtgttg atgtcgatgg ggcagggttg
      541 gtaactatgg cactctttgg tcaaccggtg ggtcccaaaa cagctgatga actgaaggag
      601 ttggtatcag ccactcgctt accttttatc gtaaaaggga ttatgactgt agacgaggcc
      661 gagctggcgg cggacgcagg agccgccgct attgttgtga gtaatcatgg cggacgcatc
      721 ttagatcata cccctggagc ggccgaagtc ttgccggcca ttgcgcgtgc ggtcaaaggt
      781 cgtgtcacga tcttagccga tggtggagta cgtagtgggg cagatgtact caagcttctt
      841 gcgttaggcg ctgatgccgt gctcgttggc cgtccgctgg ttattggggc cttcggcggt
      901 ggggctgagg gcgtgaaatt tgtcttagag aaacttaccg gagaattgaa gcaagcgatg
      961 attctgactg ggtgtgcaag cttatcggac attgatgaac gtgttctgcg cccatatccg
     1021 ggcatggagt gcgcggccgc actcgagcat catcaccatc accaccacca ccaccactga
     1081 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     1141 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     1201 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     1261 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1321 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1381 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1441 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1501 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1561 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1621 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     1681 tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1741 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1801 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1861 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1921 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1981 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2041 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2101 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2161 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2221 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2281 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2341 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2401 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2461 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2521 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2581 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2641 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2701 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2761 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2821 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2881 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2941 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3001 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3061 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3121 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3181 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3241 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3301 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3361 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3421 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3481 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3541 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3601 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3661 cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3721 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     3781 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3841 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3901 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     3961 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4021 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4081 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4141 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4201 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4261 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4321 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4381 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4441 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4501 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     4561 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     4621 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     4681 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     4741 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     4801 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     4861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     4981 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5041 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5101 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5161 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5221 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5281 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5341 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5401 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     5461 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     5521 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     5581 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     5641 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     5701 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     5761 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     5821 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     5881 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     5941 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6001 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6061 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6121 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6181 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6241 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6301 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     6361 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
//
