LOCUS       sequence-136235-        5914 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 5914)
  TITLE     SGC Oxford
REFERENCE   2  (bases 1 to 5914)
  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: PARP9
FEATURES             Location/Qualifiers
     source          1..5914
                     /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             775..792
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(845..863)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      859..906
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      943..1398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1030..1049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1240..1261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1491..2306)
                     /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(1491..2306)
                     /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     2214..2233
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2298..2317
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2428..3016
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2917..2936
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3170..3187
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3202..3344
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3270..3289)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3430..3452
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3446..3637)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4446..5528)
                     /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     5490..5509
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5529..5606)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5812..5831
                     /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 atccatgaat gcaatggtcg tgaacaacct
      181 gaccctccag attgtccagg gccacattga atggcagacg gcagatgtaa ttgttaattc
      241 tgtaaaccca catgatatta cagttggacc tgtggcaaag tcaattctac aacaagcagg
      301 agttgaaatg aaatcggaat ttcttgccac aaaggctaaa cagtttcaac ggtcccagtt
      361 ggtactggtc acaaaaggat ttaacttgtt ctgtaaatat atataccatg tactgtggca
      421 ttcagaattt cctaaacctc agatattaaa acatgcaatg aaggagtgtt tggaaaaatg
      481 cattgagcaa aatataactt ccatttcctt tcctgccctt gggactggaa acatggaaat
      541 aaagaaggaa acagcagcag agattttgtt tgatgaagtt ttaacatttg ccaaagacca
      601 tgtaaaacac cagttaactg taaaatttgt gatctttcca acagatttgg agatatataa
      661 ggctttcagt tctgaaatgg caaagaggtc caagatgctg agtttgaact gacagtaaag
      721 gtggatacgg atccgaattc gagctccgtc gacaagcttg cggccgcact cgagcaccac
      781 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
      841 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      901 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
      961 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1021 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1081 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1141 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1201 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1261 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1321 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1381 tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     1501 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1561 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1621 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1681 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1741 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1801 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1861 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1921 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1981 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2041 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     2101 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     2161 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     2221 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     2281 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     2341 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2401 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2461 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2521 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2581 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2641 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2701 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2761 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2821 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2881 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2941 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3001 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3061 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3121 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3181 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3241 ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3301 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3361 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3421 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3481 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3541 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3601 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3661 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     3721 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     3781 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     3841 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     3901 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     3961 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     4021 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4081 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4141 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4201 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     4261 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     4321 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4381 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4561 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4621 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4681 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     4741 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     4801 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     4861 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     4921 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4981 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5041 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5101 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5161 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5221 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5281 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5341 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5401 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5461 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5521 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5581 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5641 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     5701 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     5761 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     5821 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     5881 ggcgtagagg atcgagatct cgatcccgcg aaat
//
