LOCUS       sequence-354281-        6593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier U6EB70.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6593)
  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 6593)
  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: U6EB70
FEATURES             Location/Qualifiers
     source          1..6593
                     /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             1313..1342
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1395..1413)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1409..1456
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1493..1948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1580..1599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1790..1811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1974..2078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2079..2147,2148..2939)
                     /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(2297..2316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3110..3698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3599..3618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3852..3869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3884..4023
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3949..3968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4109..4131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4125..4316)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5125..6207)
                     /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     6169..6188
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6208..6285)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6491..6510
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaactgtaag attttcattt atgacgaaga
      121 tgtgggagaa ccgcgtctga acttactccc aggcaccaaa gttgaggata ttccggattc
      181 gtatcgctgc ccggtgtgtg gcttccctaa aagctatctg aaacctttag atgagacctc
      241 aacgccggaa aaggacgaaa ttaaggattt aaattactac cgtaatcgta gccgcgaaat
      301 gctcggtgga ctttgtggcg tgtatcctgt ctgtaacggc gacccggacc atctctgtat
      361 gggtcaaaag tatggtgccc ctattggctt gggtggtgcc ggacaaggca aaacgtttga
      421 agcgaattac aaagcgttgc agcaatatcg tctgaaaatg cgcgttatca agaagcacca
      481 tgaccctgag atctcgacta aaatcttcgg caagaaaatc attgctccta tcatgggcgc
      541 acccctcagt ggagccaagc ttagcctgaa tgatgctatt tcggaagagg acctgtactg
      601 ggcgctttta aagggtgccc aagactttgg cagtattggg atggtgggga atacatcgac
      661 gagcccagat gaattaggcg cgcacatcgt tgggaagaat aaaggctggg gtatccccgt
      721 gtttaagccc catccacaga agcgccttat cgagttattc aaaatcgcgg aaaagttgga
      781 cgtcattgca attggcgtag acttagatgg ttcaggtagc acctactgga caccggagaa
      841 gaacgtatac cgcaagtcag aaaatgaact gcaggaattg gtcgactgta ccaagaagcc
      901 agttttcttc aaaggcatca tgtcacctga agatgccgtt aaagtggttg actccggagc
      961 atcagcgtgt tatgttagca atcatggtgg gcgtgtcctc gacagcgggc agggagtcgc
     1021 tgatgtgctt ccggacattg cagaagagat cagcgggaag atcccaattc tggccgatgg
     1081 taccgtgcgt acgggttttg atgttttaaa agttattgcg ttaggtgcgg atgttgctct
     1141 gattggtcgt agtctggcac atgtgttaat cagtggtgga gtagccgttc agatgtacct
     1201 ggattatgta cgtgatgatt tacgtcgtgc catgattctg acaggatgca acacgcttaa
     1261 ggacgcaacg atgaatattc tgaccaaaca gaactatgcg gccgcactcg agcatcatca
     1321 ccatcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
     1381 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1441 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     1501 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1561 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1621 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1681 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1741 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1801 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1861 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1921 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     1981 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2041 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2101 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2161 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2221 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2281 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2341 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2401 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2461 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2521 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2581 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     2641 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2701 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2761 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2821 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2881 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2941 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3001 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3061 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3121 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3181 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3241 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3301 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3361 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3421 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3481 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3541 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3601 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3661 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3721 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3781 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3841 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3901 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     3961 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4021 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4081 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4141 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4201 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4261 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     4321 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     4381 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     4441 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     4501 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     4561 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     4621 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     4681 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     4741 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     4801 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     4861 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     4921 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     4981 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5041 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5101 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5161 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5221 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5281 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5341 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5401 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5461 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5521 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5581 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5641 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5701 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     5761 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     5821 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     5881 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     5941 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     6001 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     6061 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     6121 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6181 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6241 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6301 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6361 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6421 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6481 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6541 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aat
//
