LOCUS       sequence-316289-        6807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier D4N087.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6807)
  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 6807)
  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: D4N087
FEATURES             Location/Qualifiers
     source          1..6807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(71..89)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      85..132
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      169..624
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(256..275)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     466..487
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        651..755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(756..824,825..1616)
                     /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(974..993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1787..2375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2276..2295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2529..2546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2561..2703
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2629..2648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2789..2811
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2805..2996)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3805..4887)
                     /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     4849..4868
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4888..4965)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5171..5190
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5274..5293
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5274..5292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5293..5317
                     /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             5332..5354
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(6796..6807,1..18)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
ORIGIN
        1 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
       61 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
      121 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
      181 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      241 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      301 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      361 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
      421 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      481 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
      541 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
      601 acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      661 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      721 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      781 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      841 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      901 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      961 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1021 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1081 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1141 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1201 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1261 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     1321 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1381 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1441 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1501 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1561 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1621 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1681 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1741 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1801 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1861 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1921 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1981 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2041 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2101 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2161 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2221 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2281 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2341 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2401 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2461 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2521 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     2581 ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc
     2641 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     2701 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2761 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2821 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     2881 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     2941 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     3001 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     3061 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     3121 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     3181 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     3241 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     3301 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     3361 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     3421 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     3481 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     3541 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     3601 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     3661 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     3721 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     3781 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3841 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     3901 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     3961 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     4021 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     4081 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     4141 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     4201 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     4261 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     4321 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     4381 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     4441 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     4501 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     4561 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     4621 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     4681 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     4741 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     4801 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     4861 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     4921 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     4981 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     5041 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     5101 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     5161 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     5221 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     5281 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     5341 ctttaagaag gagatataca tatgccttat ccgtataaga agatgaccca agaggacttc
     5401 gacaaaatta tcgaaatcac tgacaacgaa agggtctggg tcggagacga aatcgcaaaa
     5461 gagtactaca aagacgagat gccagaatac ggtgttttcc cacccgaact gtatgtcgaa
     5521 gttttaaata aagaagaggt ctctgaaatc atgaaatacg cttatgagca aaatattccg
     5581 gttgtctgca ggggcgcagg aaccggatta gccggtggtg ctacatgcaa gtacggcggc
     5641 ataatgctgt ctgtcatgcg aatgaacaaa atatttccag tagaccataa gaatcagact
     5701 atcactgcac aaccaggtgc attgttgatc gacattcaag ccgcagcaaa ggaagagggt
     5761 ctcttctatc ccccagatcc cggtgagaag actgcttcaa ttggcggaaa tgtcattact
     5821 aatgccggtg gtatgaaggc tgttcgatat ggtctgacta gggacttcgt cagatgcatc
     5881 gaggcagtca tgcctgatgg atcaattatg aacttctcta gtaatgtagt aaagaacact
     5941 accggatttg acgtcaagga cttagtcatt ggttctgagg ggattctatg cattttgacg
     6001 gaagtcacgt taaagttgct tccagcccca acttgtagca gtacattggt aatgcctttc
     6061 agatcattgg aagaatgcgc cgatatggtc ccaaaggtgt tggacttgcc ttttgttcca
     6121 accgctatag agttcctcga aagagagctt atcgacattg tcgaaagatc actccacaaa
     6181 cttttcccag ttaagcatgg cgaagctgtc ctgatcgtta tgtatgatgc tagttctaag
     6241 gaagagttag atagggctgt ggaagcagca gctgaagcag cccttgagaa cggcgcacta
     6301 gattgtaaca ttgctggaac acctgaaaga gcagcttctg tgtgggaagt gaggggagca
     6361 attcttgagg ggatgaaagc tgacagtgtc gcccaagaag agtgcgatgt tgtcgttcca
     6421 cgatctgaga ttgctgaata cgtgaaacaa gctaagaaaa tcgcatccgc tcatggtatt
     6481 agagtagaac catgcggtca ttgcggtgat ggtaatattc atacagagat gttaagaggt
     6541 ccagaaatgt ctgacgagga gtggaagaag gcaactcacg agtctttgac tgagttatac
     6601 gcactatcaa agaagttggg tggacagcta tctggtgagc acggaattgg aaatggcagg
     6661 ttggatttct tagaggaatt tgcaggaccc aggatgatag aactgtacaa gagtattaag
     6721 agagctttcg atgataagtt gatcttgaat cccggaaaga tgatcaaaat cgatgaaaat
     6781 gctgcagctt tagaacacca ccatcac
//
