LOCUS       sequence-354362-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier D1A2X8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  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 6563)
  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: D1A2X8
FEATURES             Location/Qualifiers
     source          1..6563
                     /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             1283..1312
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1365..1383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1379..1426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1463..1918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1550..1569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1760..1781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1944..2048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2049..2117,2118..2909)
                     /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(2267..2286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3080..3668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3569..3588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3822..3839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3854..3993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3919..3938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4079..4101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4095..4286)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5095..6177)
                     /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     6139..6158
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6178..6255)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6461..6480
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gccgttgaag aatccttggt ttgaaaccgt
      121 agcagaagca caacgtcgcg ccaagaaacg tttaccttac atggtatacg gcgcattgct
      181 tgcaggcagt gagcgtggcc gtactgttca agataatacg gatgccttcg gtgaactggg
      241 ctttgcgccg cgtgtggtag gtcatcatgc gcagcgcgac ctgtcgacca ccgtcatggg
      301 agtacccacg tcaatgcccg tggtaatcag cccgactggc gtgcaagcag tccatcctga
      361 tggtgaagtc gcagtggcac gtgcggccgc gaatcgcggt gttattatgg gcctgagcag
      421 ttttgcatca aagcctattg aacaagttgt ggaagcaaac cctaatgtct tcttccagat
      481 gtattggagt ggcgatcgtg atgccattct gcaacgtatg gagcgtgcgc gtaatgcggg
      541 tgctaaaggc ctcattgtga ccctggattg gtcatttagc atgggtcgcg attggggctc
      601 cccgaccatc ccggagaaat tagatgtcaa aacaatggta aaattagccc ctgttgccct
      661 ttcccgccca ggctggctgt ggcgtttctt gaaggcccgt acattaccgg acctgaccgt
      721 tcctaatctc caaccgcccg gcggccaggc accgaccttc tttggtgcat acggtgaatg
      781 gatgcagacc ccgccaccga cgtgggatga cgtgaaatgg ctgcgtgaag aatggggcgg
      841 tccgtttatg ttaaaaggcg tgacacgtgt cgatgatgct aaacgtgcgg tcgatattgg
      901 tgtgactgcg ttaagtgttt caaatcatgg cggaaataat ttggatacca ccccagccac
      961 tattcgttta ttacccgcaa ttgcggaagc cgttggcgat caagttgaag tcctgctgga
     1021 cggtggtgtt cgtcgcggtg gtgatgtcgc gaaagcgctg gcacttggag cccgtgcagt
     1081 tttaatcggc cgtgcttacc tttggggttt ggcggccaac ggccaagcgg gtgtggaaaa
     1141 tgtactggat attctgcgct ctggtcttga ttccgctgtc cttggcctgg gtcgcagcag
     1201 tgtgcatgaa ctggccccag acgaccttgt tattccgcca gggtttcgct tagcaccggg
     1261 cgcaagcgcg gccgcactcg agcatcatca ccatcaccac caccaccacc actgagatcc
     1321 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1381 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1441 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1621 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1681 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1741 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1801 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1861 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1921 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1981 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2041 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2101 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2161 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2221 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2281 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2341 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2401 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2461 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2521 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2581 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2641 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2701 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2761 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2821 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2881 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2941 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3001 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3061 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3841 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3901 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3961 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4021 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4081 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     4141 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     4201 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     4261 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     4321 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     4381 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     4441 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     4501 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     4561 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4621 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4681 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     4741 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     4801 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     4861 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     4921 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     4981 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     5041 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     5101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5161 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     5221 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     5281 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     5341 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     5401 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     5461 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     5521 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     5581 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     5641 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     5701 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     5761 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     5821 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     5881 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5941 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     6001 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     6061 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     6121 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     6181 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     6241 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     6301 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     6361 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6421 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6481 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     6541 tcgagatctc gatcccgcga aat
//
