LOCUS       sequence-354296-        6776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier F9UTP6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6776)
  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 6776)
  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: F9UTP6
FEATURES             Location/Qualifiers
     source          1..6776
                     /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             1496..1525
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1578..1596)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1592..1639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1676..2131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1763..1782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1973..1994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2157..2261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2262..2330,2331..3122)
                     /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(2480..2499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3293..3881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3782..3801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4035..4052
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4067..4206
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4132..4151)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4292..4314
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4308..4499)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5308..6390)
                     /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     6352..6371
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6391..6468)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6674..6693
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gactgtcttc cgtgcgtttt cgaaccaaga
      121 ggtgttagat tatctggcga cccaagtcac cgacggtcag gtgtttaccg acgaggaaac
      181 actgaagaag tactctttct ccccacgcct taccgacgac gacagcggcc tggcgttggc
      241 atacgtggaa gcgcatacca caaaggacat ccaaggaacg ttaaagaccg cccgcaagta
      301 ccacatcgct gtcattcccc aggaccagac caccagtacc gtcattggcg cagatggcct
      361 gacggactct ctcattctga gcaccgctaa aatgaatcgc atcatcgaga tttcaaaggc
      421 agacagtatt gccgttgttg agcccggtgt cgtcaatggc gatcttgaca aagaggcccg
      481 taaacaaggc atgttttacg cgcccgaccc gggctcgaag ccgatctccg gtatcggcgg
      541 taatgtggca actaatgcag gcggcatgag taccgtgaag tatggcgcta ccaaagacaa
      601 cgtgttgggc gtgaaggtgg tgctggcaga tggtcgcgaa gtgaaactgg gcggtcgtac
      661 tctgaagcag gccttcggtt atgatttgac ccaattgatg attggatccg agggtaccct
      721 cggtatcatc accgaggtta tcgtgaaatt gctgccgatc cctctgggta cccctgtcat
      781 gggtgttgca ttctttgata acatgaccgc actcgcgaag gcagttactg ccatccgtat
      841 tagcggcgtt tatccaacaa tgctggaagc tctggacggt aacacagtgg ccgctttgga
      901 ccgctacgag aaaactcact acgctaagaa tagcgcggcg atgctcatct ttaagctgga
      961 caacggcggg caggacagca tggcggtggt taagaagatt cttgcagagc agcaagcaaa
     1021 cgacgtgaag attacgacgg agccgaaaga gcaagcagcg cttgagcaat tgcgccgtga
     1081 tatgctgccg gctgtgttcg ccggtcaaaa tcatattatg gaagacatgg ccgtgcccct
     1141 gtcgcagtta gcgcccttga tggactatat ccaagagtta tcgaaacgcc ttgatgtgga
     1201 aatttacacc gccggccacg ccggagatgg gaacgttcat ccgactcttg tatggccgac
     1261 cagtgtgact gaagtaccgg aaaagattat catcgccctg caagaaatgt ttaagaaaac
     1321 tctggccctg ggcgggacta tctctgggga gcacgccgtt ggcatgttga agaaccagtg
     1381 gaataatgcc gagcttggcg aggatgtcga tatgctgcag caccaaatca aagccctgtt
     1441 cgatccaatg aatatcctga acccgaaacg caagatcaat gcggccgcac tcgagcatca
     1501 tcaccatcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     1561 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1621 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1681 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1741 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1801 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1861 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1921 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1981 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2041 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2101 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     2161 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2221 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2281 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2341 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2401 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2461 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2521 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2581 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2641 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2701 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2761 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2821 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2881 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2941 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3001 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3061 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3121 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3181 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3241 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3301 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3361 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3421 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3481 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3541 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3601 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3661 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3721 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3781 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3841 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3901 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3961 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4021 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4081 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     4141 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     4201 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4261 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4321 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4381 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4441 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4501 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     4561 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     4621 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     4681 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     4741 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     4801 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     4861 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     4921 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     4981 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     5041 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     5101 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     5161 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     5221 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     5281 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5341 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5401 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5461 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     5521 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     5581 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     5641 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     5701 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     5761 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5821 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5881 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5941 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     6001 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     6061 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     6121 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     6181 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     6241 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6301 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6361 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6421 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6481 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     6541 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6601 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6661 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6721 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
