LOCUS       sequence-348071-        6416 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 6416)
  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 6416)
  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..6416
                     /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             1136..1165
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1218..1236)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1232..1279
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1316..1771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1403..1422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1613..1634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1797..1901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1902..1970,1971..2762)
                     /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(2120..2139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2933..3521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3422..3441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3675..3692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3707..3846
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3772..3791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3932..3954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3948..4139)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4948..6030)
                     /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     5992..6011
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6031..6108)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6314..6333
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaactggaaa gagttacgcg aggccgcacg
      121 cgaaaagttg aagggctatt gccgtgtctg cccctcatgt gacgggcgcg cgtgtgctgg
      181 agaagttcca ggaatgggtg gaacaggaac gggcgcgtct tttaaggcta acatcgaggc
      241 attggcacgc tatcgcttaa acttacgtac attacacggc gcaaagaacc cagacaccag
      301 cttaaccctt ttcgggaaag aactctctac tccaatctta gctgcaccca tgaccggtgt
      361 cacgtataat atgggcggag ctcttactga gcgtgaattt attggcatga tcatggcagg
      421 tgcacgccaa gcgggtacgc tcggtatgtc tggagatggc ggcgatccgg aatattataa
      481 ttcgggcctc gaagcaatcg ctgctgaagg tggccacggt attcccgtga ttaagccacg
      541 cgaacaacaa gccgtgatcg aacgcatccg ccgcgccgaa gaagccggag ccccagcggt
      601 cggtgttgat gtcgatgggg cagggttggt aactatggca ctctttggtc aaccggtggg
      661 tcccaaaaca gctgatgaac tgaaggagtt ggtatcagcc actcgcttac cttttatcgt
      721 aaaagggatt atgactgtag acgaggccga gctggcggcg gacgcaggag ccgccgctat
      781 tgttgtgagt aatcatggcg gacgcatctt agatcatacc cctggagcgg ccgaagtctt
      841 gccggccatt gcgcgtgcgg tcaaaggtcg tgtcacgatc ttagccgatg gtggagtacg
      901 tagtggggca gatgtactca agcttcttgc gttaggcgct gatgccgtgc tcgttggccg
      961 tccgctggtt attggggcct tcggcggtgg ggctgagggc gtgaaatttg tcttagagaa
     1021 acttaccgga gaattgaagc aagcgatgat tctgactggg tgtgcaagct tatcggacat
     1081 tgatgaacgt gttctgcgcc catatccggg catggagtgc gcggccgcac tcgagcatca
     1141 tcaccatcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     1201 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1261 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1321 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1381 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1441 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1501 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1561 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1621 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1681 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1741 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1801 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1861 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1921 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1981 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2041 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2101 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2161 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2221 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2281 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2341 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2401 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2461 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2521 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2581 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2641 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2701 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2761 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3061 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3601 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3661 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3721 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3781 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3841 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3901 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3961 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4021 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4081 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4141 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     4201 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     4261 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     4321 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     4381 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     4441 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     4501 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     4561 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     4621 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     4681 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     4741 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     4801 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     4861 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     4921 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4981 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5041 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5101 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     5161 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     5221 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     5281 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     5341 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     5401 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5461 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5521 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5581 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5641 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     5701 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5761 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5821 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5881 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     5941 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6001 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6061 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6121 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     6181 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6241 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6301 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6361 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
