LOCUS       sequence-337299-        6716 bp    DNA     circular SYN 11-DEC-2025
DEFINITION  Inducible expression of UniProt identifier V6AP96.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  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 6716)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 11, 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: V6AP96
FEATURES             Location/Qualifiers
     source          1..6716
                     /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             1436..1465
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1518..1536)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1532..1579
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1616..2071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1703..1722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1913..1934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2097..2201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2202..2270,2271..3062)
                     /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(2420..2439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3233..3821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3722..3741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3975..3992
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4007..4146
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4072..4091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4232..4254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4248..4439)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5248..6330)
                     /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     6292..6311
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6331..6408)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6614..6633
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gactcgtaac tttgactctt ttggaaggat
      121 tgagcgacat gaaagagcct caagatcctt cgagaaggtt agagctgggt tcgccgcttc
      181 agctgctgaa gagacttcac ttttggcctt tggaaacggt tgttcttatg gggactcttg
      241 tcacaatgat gctggtcttc ttgtacctat gagaagcgtc gctgccattg aatctttcga
      301 ccctgctacc gggattttgc aggcccagac cggagttctc ttgtctgaga ttatcgggtt
      361 cgtcgcacct catggttatt tccttccggt taccccagga accaggttcg ttacattggg
      421 cggagccatc gccaacgatg ttcatgggaa gaatcatcac ctgcgaggta cttttggctg
      481 ccacgtagag tctttggagc tcctacgttc cgacggtgtg cagtaccact gtagtcttca
      541 cgagaacgag gggctctttg cagcaaccat aggtggaatg ggtttaacag gagtaattgt
      601 gagcgcaagg ttgagattaa tgcccgtagg atccctagat gttgaagaga gaattgtccc
      661 tttcgataac ttggacgcat attttgatat ggctgaggcc gctgaccgtc aaaacgaata
      721 tgctgttgct tgggttgatc agttgagtcg tgggaaaact gcaggtcgtg gagtactaat
      781 aaccggtaat cacgccggga acggcaatcg ttctgtggaa gacgagcgtg tgagactggg
      841 tgtccctttc gacctgccat tttcagtttt gaatcgtgtt tcattaagac tattcaacat
      901 ggcctacttt aatgccaaga gacgtaagca gaagccacat ttggcttcat ttggatcctt
      961 cttctatcca ttagacggag ttgggcagtg gaatagattg tacgggccag ctggcctgta
     1021 tcagcatcaa agtgttattc cctttgatgc tgcaagacac atcattcccg ctatgctagc
     1081 tgctagtaga gaggccggtc aggcttcctt cctaactgtc ttgaagagat tcggtgatac
     1141 ggcatcccca ggtttgttgt ctttcccaaa gcctggttac acactcacta tggacttccc
     1201 taaccgaggt ggtaagacac gtgcacttct gcagactttg gataacatga ctatcggtgc
     1261 tggtggccgt gttaatcctt ataaagacca aagaatggga gcagctacct tcaaagctgg
     1321 ttttcctcaa tggccagagt tggaacagtg gagagataag agattctgct cagacttttg
     1381 gagacgaact gctctggccc cagaattgca gcaaggacga gctgctgctt tagaacacca
     1441 ccatcaccac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     1501 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1561 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1621 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1681 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1741 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1801 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1861 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1921 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1981 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2041 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     2101 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2161 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2221 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2281 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2341 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2401 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2461 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2521 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2581 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2641 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2701 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2761 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2821 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccagctggct
     2881 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2941 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3001 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3061 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3121 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3181 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3241 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3301 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3361 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3421 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3481 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3541 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3601 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3661 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3721 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3781 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3841 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3901 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3961 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4021 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     4081 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     4141 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4201 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4261 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4321 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4381 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4441 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     4501 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     4561 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     4621 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     4681 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     4741 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     4801 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     4861 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     4921 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     4981 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     5041 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     5101 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     5161 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     5221 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5281 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5341 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5401 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     5461 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcat cgtatcccac
     5521 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     5581 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     5641 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     5701 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5761 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5821 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5881 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5941 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     6001 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     6061 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     6121 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     6181 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6241 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6301 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6361 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6421 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     6481 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6541 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6601 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6661 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
