LOCUS       sequence-224811-        5527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Strep-tagged murine SAN1 nuclease in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5527)
  AUTHORS   Andrews AM, McCartney HJ, Errington TM, D'Andrea AD, Macara IG
  TITLE     A senataxin-associated exonuclease SAN1 is required for resistance 
            to DNA interstrand cross-links.
  JOURNAL   Nat Commun. 2018 Jul 3;9(1):2592. doi: 10.1038/s41467-018-05008-8.
  PUBMED    29968717
REFERENCE   2  (bases 1 to 5527)
  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: "Nat 
            Commun."; date: "2018-07-3"; volume: "9(1):2592. doi"; pages: " 
            10.1038/s41467-018-05008-8"
COMMENT     Sequence Label: pASK-mSAN1-Streptag
FEATURES             Location/Qualifiers
     source          1..5527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..56
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7..25
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             95..117
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2489..2512
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      2603..3058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2690..2709)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2900..2921
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3085..3189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3190..3258,3259..4050)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3408..3427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4057..4683
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4133..4152)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     rep_origin      4836..5424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5325..5344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttgacactc tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt
       61 gaaatgaata gttcgacaaa aatctagaaa taattttgtt taactttaag aaggagatat
      121 acaaatgggt gtaagaggtt tgcaggggtt tgtggggagt acgtgcccac acatatgtac
      181 catagtcaat atccacgagc tggcagagcg tcaccgaaac aagtaccctg gatgtacgcc
      241 taccatcgtg gtggacgcca tgtgctgtct ccgatactgg tacacagcag agtcgtgggt
      301 ctgcggcggc cagtggagag aatactactg tgctttgaga aattttgttg cagccttcac
      361 ctctgccggc atcaagttga tattcttctt tgatggcatg gtggagccgg ggaagcgaga
      421 tgagtgggta aagcgtagac tcaaaaacaa ccgggagata tccaagattt ttcactacat
      481 caagtcaaaa agagatcagc caggcagaaa catgttcttc attccctctg ggctggccat
      541 attcacacgg ttcgccctga agacactggg ccaggagact ttctgttcat tacaggaggc
      601 agactacgag gtggcttctt atggcctcca gcacaactgt ctggggatcc ttggggaaga
      661 cactgactac ttaatttatg acacttgccc ctacttttca attggcgatc tctgcctaga
      721 gagtctccaa accatcatgc tctgtcgaga gaagctctgt gagagcctgg gcctccgtgt
      781 ggcagacctt cctcttctgg cctgtctgct tggcaatgac atcactccag agagcatgtt
      841 tgaaagcttt cggtacaagt gcttgtcatc ctatgcttct gtgaaagaga atgctggcaa
      901 aaaaggaaat attatcctag ctgtctcaga ctatatctcc aaagttcttc atttgtatca
      961 aggtgagaag aagatagaag agatgctacc cttggggcca aacaaagctc tcttttataa
     1021 aggagtaacc tcatatcttt tgccagggca gaaatctcca tggttagttc aaaaacccaa
     1081 aggcatgata acagacaaac agcaaatggt gagtttaaat cccgaatcca agcaagaagt
     1141 ccctatgtgt attgatcctg aattcaagca agaagttcct gtgtgcacta atcccgagtc
     1201 catgcaagaa gttcccatgt gtatggatcc tgagcccaac caggaagctt ccatgtgcac
     1261 agatcccgaa tccaaacaag aagttcccat gtgcacagac tccgaatcca agccagaagt
     1321 ttcccaatat acaaacccag agagcaagca aaaattacct tcaggaatag acactgaatt
     1381 taacctagaa gcactcatgt gtacacaccc tgaatttaaa caagaagatg tcatggatat
     1441 ggaacctgaa ataaagcaag taaccatggt ttccgagtct gagatcttaa aggttgccag
     1501 gatgcatcac gtccactcag aaagctacct agtgtacaac attctgagca gtggggagat
     1561 agaatgcagt aacaccctgg aggatgagct ggaccaagcc ctcccgagcc aagcctttat
     1621 ctaccgccca gtccggcagc gcgtctacgc actcctactc ggggactgga aagatggagc
     1681 cagcactggc ccagtggtca aggagtggtt tgtgtacccc ggtaactccc tgaagcaccc
     1741 agacctggtt aggcctctgc agatgaccat tcaaggggga acacctagct tggaagttct
     1801 gtggctgagc caagagccag cagtgcaggc ccagcgcctg gacaccttgc tagcctgctt
     1861 caacctttcc tcctcaagag aagagctgca agcagttgag agcccgttga gagcgctgtg
     1921 ctgcctcctg atctacctct ttgtccaggt ggacactctt tccctggaag atctgcatgc
     1981 atttatcgct caggccttgt gcctccaagg aaaatcaacc tcccagctca tgcacttaca
     2041 gctggattac attaactcca gagctgtgca gctgggatcc ctcctcgtcc gtggcctcac
     2101 cacattggta ctagtcaata gcgcatgtgg cttcccctgg acaacaagtg agtttatgcc
     2161 ctggaatgtg tttgatggca agcttttcca tcagaagtac ctgcagtctg aaaagggata
     2221 tgccgtggaa gttcttttgg aacaaaatag atcgtggctc accaagttcc acaacctgaa
     2281 ggcagtggtc tgcaaggcct gctctaagga gaaccggcgc atcgtaggca gaacgcactg
     2341 ggactctcct tacacaggga ggcaagggag acagggctac agctcctaca ggacagactc
     2401 cacacatggc cattctgggc agtcatggag aaaccagggt tcagggggca gacagcatga
     2461 gcgtaaccac tggagaagat acagcgcttg gagccacccg cagttcgaaa aataataagc
     2521 ttgacctgtg aagtgaaaaa tggcgcacat tgtgcgacat tttttttgtc tgccgtttac
     2581 cgctactgcg tcacggatct ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2641 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2701 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2761 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2821 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2881 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2941 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3001 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttca
     3061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3781 actctagctt cccggcaaca attgatagac tggatggagg cggataaagt tgcaggacca
     3841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3901 cgtggctctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4021 ataggtgcct cactgattaa gcattggtag gaattaatga tgtctcgttt agataaaagt
     4081 aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc
     4141 cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat
     4201 aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt
     4261 tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga
     4321 tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa
     4381 aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta
     4441 gagaatgcat tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa
     4501 gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg
     4561 ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta
     4621 ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct
     4681 taaaagcagc ataacctttt tccgtgatgg taacttcact agtttaaaag gatctaggtg
     4741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4981 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5461 ttttgctggc cttttgctca catgacccga caccatcgaa tggccagatg attaattcct
     5521 aattttt
//
