LOCUS       sequence-354450-        6608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier C4VMW0.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6608)
  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 6608)
  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: C4VMW0
FEATURES             Location/Qualifiers
     source          1..6608
                     /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             1328..1357
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1410..1428)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1424..1471
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1508..1963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1595..1614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1805..1826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1989..2093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2094..2162,2163..2954)
                     /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(2312..2331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3125..3713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3614..3633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3867..3884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3899..4038
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3964..3983)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4124..4146
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4140..4331)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5140..6222)
                     /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     6184..6203
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6223..6300)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6506..6525
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gacagtctac tacaagggat ttcctcagag
      121 tgacagaaat gaagctatca agatggttaa tgttgatgag ttggaagaca gggtgaggaa
      181 ggttatgcca gaagctgcat actactatat tgcttctgga tctgagaacg aatggacttg
      241 gcgaaataac actgccgcct tcaatcactt ccagattgtc ccacgaagct taactaacat
      301 ggacaacccg tctactgaaa cccaatttat gggtatggac ctaaagacac ccattatgat
      361 ctgtccaatt gcctgtcacg gaatcgccca taaagatgct gaagtcgcaa ccgcacaggg
      421 agctaaggct gccggagctt tgttttcgtc gagcacttac gctaatagga gtgtagaaga
      481 tatagctact gctactggag attccccaaa attcttccaa ctctatttgt ctaaggactg
      541 ggacttcaat aagatggtgt tcgacgcagt gaaaagtgct ggctacaaag gaatcatgct
      601 tactgttgat gcattagtat ctggttatcg tgaggcaaac ctgcgaacta actttacttt
      661 ccctgttcca ttagatttct tcacccgata cgtcggtgcc gagggagaag gaatgtctgt
      721 cgcccaaatg tatgctaaca gtgcccaaaa gatcggacca gctgatgtag ccaaaattaa
      781 agaaatgtcg ggcttacccg tattcgtcaa aggagtgatg aatgccgagg acgcatatat
      841 ggccattggc gcaggcgctg atggaatagt cgtctccaat catggtggga gagaaatcga
      901 cacggcacca gctaccatcg acatgttacc tgagatcgca gctgcagtaa atggcagggt
      961 tccaatcatt ctggactctg gtgtcaggcg aggatctcac gtatttaagg cacttgcttt
     1021 aggagcagat ttggtcggta tcggtagacc tttcctgtac ggtcttgctt tgggcggtgc
     1081 caagggagtg gagtccgtca tcaatcagat caacaacgaa ttcaaaattt tgatgcaact
     1141 aactggctgt aagaccgtag aggacgtcaa acacgcagac atcagacaaa ttaactatac
     1201 agcagacaac ttgcctagta ataccgaccc ttctgtgaga agagcctatc ctgttactaa
     1261 ggaaaaccaa atggaaggta cacaagatgc tgctaccggg gcttctaagc acgcagcagc
     1321 cttagaacac caccatcacc accaccacca ccaccactga gatccggctg ctaacaaagc
     1381 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1441 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     1501 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1561 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1621 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1681 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1741 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1801 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1861 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1921 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     1981 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2041 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2101 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2161 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2221 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2281 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2341 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2401 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2461 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2521 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2581 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     2641 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2701 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2761 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2821 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2881 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2941 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3001 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3061 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3121 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3181 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3241 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3301 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3361 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3421 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3481 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3541 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3601 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3661 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3721 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3781 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3841 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3901 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     3961 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     4021 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4081 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4141 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4201 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4261 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4321 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4381 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4441 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     4501 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     4561 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     4621 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     4681 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     4741 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
     4801 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4861 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4921 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4981 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     5041 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     5101 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5161 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5221 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     5281 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     5341 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5401 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5461 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5521 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5581 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5641 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5701 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5761 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5821 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5881 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     5941 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     6001 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     6061 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     6121 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     6181 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     6241 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     6301 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6361 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     6421 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6481 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6541 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     6601 cgcgaaat
//
