LOCUS       sequence-220935-        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        36..91
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    42..60
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    72..90
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2524..2547
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      2638..3093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2725..2744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2935..2956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3120..3224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3225..3293,3294..4085)
                     /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(3443..3462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4092..4718
                     /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(4168..4187)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     rep_origin      4871..5459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5360..5379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatcgaatg gccagatgat taattcctaa tttttgttga cactctatca ttgatagagt
       61 tattttacca ctccctatca gtgatagaga aaagtgaaat gaatagttcg acaaaaatct
      121 agaaataatt ttgtttaact ttaagaagga gatatacaaa tgggtgtaag aggtttgcag
      181 gggtttgtgg ggagtacgtg cccacacata tgtaccatag tcaatatcca cgagctggca
      241 gagcgtcacc gaaacaagta ccctggatgt acgcctacca tcgtggtgga cgccatgtgc
      301 tgtctccgat actggtacac agcagagtcg tgggtctgcg gcggccagtg gagagaatac
      361 tactgtgctt tgagaaattt tgttgcagcc ttcacctctg ccggcatcaa gttgatattc
      421 ttctttgatg gcatggtgga gccggggaag cgagatgagt gggtaaagcg tagactcaaa
      481 aacaaccggg agatatccaa gatttttcac tacatcaagt caaaaagaga tcagccaggc
      541 agaaacatgt tcttcattcc ctctgggctg gccatattca cacggttcgc cctgaagaca
      601 ctgggccagg agactttctg ttcattacag gaggcagact acgaggtggc ttcttatggc
      661 ctccagcaca actgtctggg gatccttggg gaagacactg actacttaat ttatgacact
      721 tgcccctact tttcaattgg cgatctctgc ctagagagtc tccaaaccat catgctctgt
      781 cgagagaagc tctgtgagag cctgggcctc cgtgtggcag accttcctct tctggcctgt
      841 ctgcttggca atgacatcac tccagagagc atgtttgaaa gctttcggta caagtgcttg
      901 tcatcctatg cttctgtgaa agagaatgct ggcaaaaaag gaaatattat cctagctgtc
      961 tcagactata tctccaaagt tcttcatttg tatcaaggtg agaagaagat agaagagatg
     1021 ctacccttgg ggccaaacaa agctctcttt tataaaggag taacctcata tcttttgcca
     1081 gggcagaaat ctccatggtt agttcaaaaa cccaaaggca tgataacaga caaacagcaa
     1141 atggtgagtt taaatcccga atccaagcaa gaagtcccta tgtgtattga tcctgaattc
     1201 aagcaagaag ttcctgtgtg cactaatccc gagtccatgc aagaagttcc catgtgtatg
     1261 gatcctgagc ccaaccagga agcttccatg tgcacagatc ccgaatccaa acaagaagtt
     1321 cccatgtgca cagactccga atccaagcca gaagtttccc aatatacaaa cccagagagc
     1381 aagcaaaaat taccttcagg aatagacact gaatttaacc tagaagcact catgtgtaca
     1441 caccctgaat ttaaacaaga agatgtcatg gatatggaac ctgaaataaa gcaagtaacc
     1501 atggtttccg agtctgagat cttaaaggtt gccaggatgc atcacgtcca ctcagaaagc
     1561 tacctagtgt acaacattct gagcagtggg gagatagaat gcagtaacac cctggaggat
     1621 gagctggacc aagccctccc gagccaagcc tttatctacc gcccagtccg gcagcgcgtc
     1681 tacgcactcc tactcgggga ctggaaagat ggagccagca ctggcccagt ggtcaaggag
     1741 tggtttgtgt accccggtaa ctccctgaag cacccagacc tggttaggcc tctgcagatg
     1801 accattcaag ggggaacacc tagcttggaa gttctgtggc tgagccaaga gccagcagtg
     1861 caggcccagc gcctggacac cttgctagcc tgcttcaacc tttcctcctc aagagaagag
     1921 ctgcaagcag ttgagagccc gttgagagcg ctgtgctgcc tcctgatcta cctctttgtc
     1981 caggtggaca ctctttccct ggaagatctg catgcattta tcgctcaggc cttgtgcctc
     2041 caaggaaaat caacctccca gctcatgcac ttacagctgg attacattaa ctccagagct
     2101 gtgcagctgg gatccctcct cgtccgtggc ctcaccacat tggtactagt caatagcgca
     2161 tgtggcttcc cctggacaac aagtgagttt atgccctgga atgtgtttga tggcaagctt
     2221 ttccatcaga agtacctgca gtctgaaaag ggatatgccg tggaagttct tttggaacaa
     2281 aatagatcgt ggctcaccaa gttccacaac ctgaaggcag tggtctgcaa ggcctgctct
     2341 aaggagaacc ggcgcatcgt aggcagaacg cactgggact ctccttacac agggaggcaa
     2401 gggagacagg gctacagctc ctacaggaca gactccacac atggccattc tgggcagtca
     2461 tggagaaacc agggttcagg gggcagacag catgagcgta accactggag aagatacagc
     2521 gcttggagcc acccgcagtt cgaaaaataa taagcttgac ctgtgaagtg aaaaatggcg
     2581 cacattgtgc gacatttttt ttgtctgccg tttaccgcta ctgcgtcacg gatctccacg
     2641 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2701 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2761 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2821 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2881 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2941 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3001 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3061 cgaattttaa caaaatatta acgcttacaa tttcaggtgg cacttttcgg ggaaatgtgc
     3121 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3181 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3241 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3301 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3361 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3421 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3481 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3541 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3601 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3661 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3721 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3781 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattga
     3841 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3901 gctggtttat tgctgataaa tctggagccg gtgagcgtgg ctctcgcggt atcattgcag
     3961 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4021 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4081 ggtaggaatt aatgatgtct cgtttagata aaagtaaagt gattaacagc gcattagagc
     4141 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
     4201 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
     4261 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
     4321 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
     4381 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
     4441 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
     4501 cagtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
     4561 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
     4621 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
     4681 gattagaaaa acaacttaaa tgtgaaagtg ggtcttaaaa gcagcataac ctttttccgt
     4741 gatggtaact tcactagttt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4801 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4861 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4921 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4981 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5041 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5101 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5161 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5221 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5281 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5341 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5401 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5461 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatga
     5521 cccgaca
//
