LOCUS       sequence-354301-        6791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0D6I3R9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6791)
  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 6791)
  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: A0A0D6I3R9
FEATURES             Location/Qualifiers
     source          1..6791
                     /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             1511..1540
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1593..1611)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1607..1654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1691..2146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1778..1797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1988..2009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2172..2276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2277..2345,2346..3137)
                     /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(2495..2514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3308..3896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3797..3816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4050..4067
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4082..4221
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4147..4166)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4307..4329
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4323..4514)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5323..6405)
                     /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     6367..6386
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6406..6483)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6689..6708
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaccgaaccg ctcgctacaa agaccgcgat
      121 cttcgatgtt gcagtgattg gtgccggtgt tgtgggctgc gcgatggcgc gccgttttgc
      181 attagaaggt gctcgcgttg tattgctgga aaagtcgacc gatattttat ctggcgccag
      241 taaaggtaac agtgcgattt tacatactgg ctttgatgct ccgccaggaa gcattgaact
      301 ggcctgcatg caagaaggat accgcgaata tcgcgatatc cacgcccgct tgaatttgcc
      361 tctcctggaa acaggggcct tagtggcagc gtggagtgat gaagatctcg cgcgcttgcc
      421 agcgatcgta gagcaagccc atggtaatgg ggtagatgac gtgcgccagg ttggtcgcga
      481 tgaggtttta gcgttagagc cacaattagc gggcaacgtg ttaggcgccg tgcgcgtccc
      541 tggtgaacat ttggtggatc cctggtcggc gccgttagcg tacctgcagc aagccttggc
      601 acatggggcc gaagcatgtt ttgacgttga ggttctggac ggtgcatttg acggcggtga
      661 gtggctgctg cacacgcgtc gcggggattt gcgcgcacgt caagtgatta attgtgcggg
      721 tctgttcgga gaccaactcg agcagcgtct tttgggcgcg gcctcttttg caattcatcc
      781 gcgtaaaggt caatttgtcg tgtttgataa agccgccgcg gcattattac gtcacatttt
      841 attgcccgtg ccgaatgagc gcactaaagg cgttgttttc acccgtacag tgtttggcaa
      901 tttattagca ggtcctacgg cggaagaaca agacgatcgt gaacaggctc gtgttgattc
      961 tgatacgctg caacgtctga tcgacgcggc tgtggaacgt atcccgggtc tccgtggtat
     1021 gccagttact gctacttacg ctggtttacg tccggccagt gagaagaagg aataccgtat
     1081 tcgccaggtg gatggtcgta attggatttc ggtgggcggc attcgtagca ccggccttac
     1141 cgccgccctg ggtattgcgc gtcacgttta tcgtctctac caaggtgaac gtttccatga
     1201 agccgtacgt gaaccgcttt ggccacaagt gccgaattta gcagaacatt tgccgcgtga
     1261 ctggcaacgc ccgggatatg atgaaattgt gtgtcactgt gagatggtga ctcgccgcga
     1321 gattctggcg gctttagaag gacctctccc agctggcgat tttggcggac tgaaacgccg
     1381 tacccgtgcc tgtatggggc gctgtcaagg tttctattgc aacgcacgcg tcgcggaatt
     1441 atctgctgga cgtctggcgc aaccactggc gaccggtagc tgtcatgagc atcacgcggc
     1501 cgcactcgag catcatcacc atcaccacca ccaccaccac tgagatccgg ctgctaacaa
     1561 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1621 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     1681 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1741 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1981 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2041 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2101 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2161 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2221 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2281 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2341 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2401 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2461 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2521 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2581 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2641 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2701 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2761 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2821 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2881 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2941 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3001 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3061 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3121 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3181 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3241 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3421 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3721 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3901 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3961 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4021 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4081 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     4141 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     4201 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4261 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4321 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     4381 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     4441 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     4501 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     4561 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     4621 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     4681 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     4741 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     4801 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     4861 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     4921 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     4981 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     5041 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     5101 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     5161 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     5221 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     5281 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5401 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     5461 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     5521 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     5581 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     5641 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     5701 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     5761 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     5821 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     5881 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     5941 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6001 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6061 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     6121 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     6181 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     6241 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     6301 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     6361 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     6421 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     6481 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     6541 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     6601 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     6661 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     6721 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     6781 tcccgcgaaa t
//
