LOCUS       sequence-193414-        6687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6687)
  TITLE     SGC Oxford 2017
REFERENCE   2  (bases 1 to 6687)
  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"
COMMENT     Sequence Label: RECQL5 (RECQL5A-c104)
FEATURES             Location/Qualifiers
     source          1..6687
                     /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             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1552..1569
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1622..1640)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1636..1683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1720..2175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1807..1826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2017..2038
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2267..3082)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2267..3082)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2990..3009
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3074..3093
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3204..3792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3693..3712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3946..3963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3978..4117
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4043..4062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4203..4225
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4219..4410)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5219..6301)
                     /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     6263..6282
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6302..6379)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6585..6604
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccatggac cctgagcggc gagtccggag
      181 tacgctgaag aaggtctttg ggtttgactc ttttaagacg cctttacagg agagtgcgac
      241 catggctgta gtaaaaggta acaaggacgt ctttgtgtgc atgcccacag gggcaggaaa
      301 atccctatgc tatcagctcc ctgctctgtt ggccaaaggc atcaccattg tagtctctcc
      361 tctcattgct ttgattcagg accaagtgga ccacttgcta accctaaagg tacgagtaag
      421 ttccctgaac tcgaagctct ctgcacagga aaggaaggag ctgcttgctg acctggagcg
      481 agaaaagccc cagaccaaga ttctgtacat caccccagag atggcagctt catcctcctt
      541 ccagcccacc ctgaactccc tggtgtcccg ccacctgctg tcttacttgg tggtggatga
      601 agctcattgt gtttcccaat gggggcatga ctttcgtcct gactacttgc gtctgggtgc
      661 cctgcgctcc cgcctgggac atgccccttg tgtggctctg accgccacag ccaccccaca
      721 ggtccaagag gacgtgtttg ctgccctgca cctgaagaaa ccagttgcca tcttcaagac
      781 tccctgcttc cgggccaacc tcttctatga tgtgcaattc aaggaactga tttctgatcc
      841 ctatgggaac ctgaaggact tctgccttaa ggctcttgga caggaggctg ataaagggtt
      901 atctggctgc ggcattgtgt actgcaggac tagagaggct tgtgaacagc tggccataga
      961 gctcagctgc aggggtgtga acgccaaggc ttaccatgca gggctgaagg cctctgaaag
     1021 aacgctggtg cagaacgact ggatggagga gaaggtccct gtaattgttg caaccattag
     1081 ttttgggatg ggagtggata aagccaatgt caggtttgtc gcccattgga atattgccaa
     1141 gtctatggct gggtactacc aggagtctgg ccgggctggc agggatggga agccttcctg
     1201 gtgccgtctc tattactcca ggaatgaccg ggaccaagtc agcttcctga tcaggaagga
     1261 agtagcaaaa ctccaggaaa agagaggaaa caaagcatct gataaagcca ctatcatggc
     1321 ctttgatgcc ctggtgacct tctgtgaaga actggggtgc cgccatgccg ccattgccaa
     1381 gtacttcggg gatgcgctgc ctgcctgcgc caaaggctgc gaccactgcc agaaccccac
     1441 ggccgtgcgg aggcggctgg aggccttgga gcgcagcagc agctggtgac agtaaaggtg
     1501 gatacggatc cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac
     1561 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1621 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1681 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     1741 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1801 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1861 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1921 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1981 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2041 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2101 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2161 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2221 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     2281 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     2341 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     2401 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     2461 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     2521 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2581 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     2641 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     2701 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     2761 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     2821 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     2881 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     2941 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     3001 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     3061 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     3121 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3841 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3901 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3961 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4021 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4081 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4141 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4201 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4261 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4321 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4381 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4441 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4501 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4561 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4621 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4681 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4741 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4801 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4861 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4921 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4981 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     5041 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     5101 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5161 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     5221 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5281 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     5341 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     5401 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     5461 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     5521 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5581 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5641 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5701 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5761 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5821 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5881 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5941 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     6001 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     6061 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     6121 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     6181 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     6241 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     6301 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     6361 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     6421 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     6481 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6541 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     6601 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     6661 aggatcgaga tctcgatccc gcgaaat
//
