LOCUS       sequence-354459-        6881 bp    DNA     circular SYN 15-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 6881)
  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 6881)
  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: V6AP96
FEATURES             Location/Qualifiers
     source          1..6881
                     /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             1601..1630
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1683..1701)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1697..1744
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1781..2236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1868..1887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2078..2099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2262..2366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2367..2435,2436..3227)
                     /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(2585..2604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3398..3986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3887..3906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4140..4157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4172..4311
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4237..4256)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4397..4419
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4413..4604)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5413..6495)
                     /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     6457..6476
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6496..6573)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6779..6798
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaacattctg tatgacgagc gcttggatgg
      121 accattacct caagttgata aagatggtct gcttgcggaa ctccgtctcc gccttccgga
      181 cttagagctg ctgcatgccg cggaagatct ccgtccgtac gagtgtgatg gcctcagcgc
      241 ctatcgctgt actccgcttt tggtagcact gcctgagcgt attgagcaag tacagggttt
      301 actcgcatta tgtcatcgcc tgaaagtgcc cgtcgtcgcc cgcggcgcgg gaactggact
      361 ttcgggtggt gcattacctc tcgagaacgg ggttttattg gttatggcgc gttttcgccg
      421 tatcctggaa attgacccac ttggccgctt tgcgcgcgta caacccgggg tacgcaatct
      481 ggcgatttcc caggctgctg ctcctcacgg tctttattat gcgcccgacc caagcagtca
      541 aatcgcttgc tccatcggtg gtaatgtggc cgaaaatgca ggtggtgtgc attgtctgaa
      601 atatggcctt accgttcaca acctgcttca agtagatatt gtgacccttg agggcgaacg
      661 tctttcatta ggctccagcg cattggattc tgcaggtttt gatttactgg cgctctttac
      721 cggctcagag ggcctgctgg gtgttgttgt agaagttaca gtacgcctgt tgccgcgtcc
      781 gccggttgcc aaggtattgt tagcctcatt cgatgacgtg gaaagcgccg gtcgtgctgt
      841 agccgatctg attggtgccg gtatcgtacc ggctggcctt gaaatgatgg ataacctcag
      901 cattcgtgcg gccgaagact tcatccatgc gggctacccg gtggacgccg cggccattct
      961 gctctgcgaa cttgacggtg tagaggcgga cgtgcatgaa gattgcgaac gcgtgcgcga
     1021 gctgtttgag gcagcgggga gcatttcagt gcgccaagcc caagacgaag ctgaacgtca
     1081 gcgtttctgg gcaggacgta agaatgcgtt tccagctgtt ggccgtattt cgccggatta
     1141 ctactgtatg gatgggagta ttccccgtcg tgaactgccg cgcgtgctgc atggcatcgc
     1201 ggaactgagc cgtgaatacg gtctgcgtgt cgccaatgtt ttccatgccg gagatggaaa
     1261 tatgcaccct ttgattctgt tcgatgccaa tctgccagga gagctggaac gtgccgaagc
     1321 attgggtggc cgtattttgg aattatgtgt cgctgttggt ggctcaatta ccggcgagca
     1381 tggcgtaggc cgcgagaaga tcaaccaaat gtgtgcccag ttcaacgccg acgaattgac
     1441 cctgttccac gctgtgaagg ccgcgtttga tccagcaggt ctgttaaacc cgggtaagaa
     1501 cgttcctact cttcatcgct gtgcggaatt tggtgctatg catgtacatc atggccgcct
     1561 gccctttccg gaacttgagc gtttcgcggc cgcactcgag catcatcacc atcaccacca
     1621 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1681 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1741 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1801 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1861 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1921 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1981 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2041 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2101 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2161 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2221 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2281 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2341 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2401 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2461 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2521 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2581 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2641 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2701 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2761 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2821 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2881 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2941 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3001 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3061 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3121 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3181 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3241 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3301 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3361 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3421 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3481 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3541 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3601 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3661 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3721 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3781 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3841 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3901 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3961 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4021 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4081 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4141 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4201 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4261 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4321 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4381 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4441 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4501 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4561 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4621 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4681 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4741 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4801 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4861 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4921 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4981 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     5041 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     5101 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5161 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5221 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5281 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5341 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5521 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     5581 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5641 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     5701 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5761 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5821 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5881 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5941 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     6001 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     6061 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     6121 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     6181 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     6241 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6301 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6361 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6421 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6481 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     6541 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6601 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6661 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6721 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6781 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6841 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
