LOCUS       sequence-354302-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0J1FVG5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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: A0A0J1FVG5
FEATURES             Location/Qualifiers
     source          1..6506
                     /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             1226..1255
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1308..1326)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1322..1369
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1406..1861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1493..1512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1703..1724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1887..1991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1992..2060,2061..2852)
                     /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(2210..2229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3023..3611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3512..3531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3765..3782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3797..3936
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3862..3881)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4022..4044
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4038..4229)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5038..6120)
                     /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     6082..6101
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6121..6198)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6404..6423
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggaacaggtg gacatcgtgg tcattggcgc
      121 tggggtcgta ggcttagcgg tcgcggccga gctttcaaag caattcaacg atcgctccct
      181 ggtgctgctg gaacgccact cgaaattcgg tcaagaaact tcgagccgta atagcgaggt
      241 gattcatgca ggcatttact atccgagcgg ctcgctgaaa gctaatctgt gcgtcgaagg
      301 aaatcaacgc ctgtatcatt tctgtcgcgc ctggaatatt ccgcatcagc gtattggaaa
      361 gctcattatt gcacgttcag agcaagaaat tccggtattg gatagtttat tagagcaagg
      421 gcagaagaat ggcgtgaatg acctggagct gattgacaac gaccaaattg cgcgtttgga
      481 acctcacatc tctgcagttg cagcggtgct ctctccttct acgggcattg ttgacagcca
      541 tcagctgatg ctgcgcctgg aatggctggc attggagcag ggtgccgtta ttgcctacca
      601 gcacaacgta cagggagtgg agcacatgga taagggttat gaggttacct accgtaaccc
      661 ggctgggcag ttagaaactc tgcgctgtca gtggctgatt aattgtgcag gcctgaattc
      721 tgatcacatt ccctcgttct tgggcatcga catcgacaat agcggctacc gtatttatcc
      781 ttgcaaaggg gagtacttta gcgtgccgtc tagtaagtcg gcgctggtaa gccacctgat
      841 ttatccgccg ccgttgaaag cgctgaaagg cttgggtatt cacgccacca agagcctgga
      901 cggtcgcctg cgcttcgggc ctaatgcgtt ctacaccgaa accctcaatt atgacgtcga
      961 tgaggcccat ggtcaggaat tttataacgc cgtgaccaca tatatgccct tccttgactc
     1021 agaggacttc ctgccggata tggcaggcat tcgtccgaag attcaagcgc cgggcgatcc
     1081 ggtccgcgac ttcattgttt gccacgaagc ggaacgtggg ttcgaaggtt taattaatct
     1141 gatcggtatc gaatcgcccg ggttgacgtc atcgttgact ctcgccgaga aagtcaaaga
     1201 cttaatgtac gcggccgcac tcgagcatca tcaccatcac caccaccacc accactgaga
     1261 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1321 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1381 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1441 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1501 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1561 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1621 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1681 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1741 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1801 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1861 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1921 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2221 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2281 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2341 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2401 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2521 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2581 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2641 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2761 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2821 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2881 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2941 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3001 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3781 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3841 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3901 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     3961 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4021 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4081 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     4141 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     4201 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     4261 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     4321 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     4381 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     4441 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     4501 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     4561 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     4621 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     4681 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     4741 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     4801 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     4861 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     4921 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     4981 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     5041 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5101 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     5161 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     5221 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     5281 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     5341 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     5401 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     5461 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     5521 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     5581 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     5641 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     5701 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     5761 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     5821 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     5881 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     5941 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     6001 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     6061 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     6121 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     6181 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     6241 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     6301 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6361 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     6421 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     6481 ggatcgagat ctcgatcccg cgaaat
//
