LOCUS       sequence-224810-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged SAN1 nuclease in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  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 7400)
  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: pK-SAN1-Flag
FEATURES             Location/Qualifiers
     source          1..7400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3647..3670
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3687..3696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3700..3834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3737..3756)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3791..3810
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3903..4260
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3903..4094
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3926..3946)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4111..4246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4280..4297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4280..4296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4289..4311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4509..4964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4596..4615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4806..4827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4981..5000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5100..5122
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5160..5178)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5246..5350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5351..5419,5420..6211)
                     /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(5569..5588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6382..6970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6871..6890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7124..7141
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7258..7279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7294..7299,7300..7317,7318..7324)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7332..7348
                     /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)."
     primer_bind     7337..7359
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7356..7372
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7356..7372
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctaa
      901 gctttctaga atgggtgtga gaggtttgca aggatttgtg ggaagtacct gcccacatat
      961 atgtacagta gtaaatttca aagaactggc agagcaccac cgaagcaagt atcctggatg
     1021 tacccctacc attgtggttg atgccatgtg ttgtctcaga tattggtata ctccagaatc
     1081 ttggatctgc ggtggccagt ggcgagaata cttttctgct ttgcgagatt ttgttaaaac
     1141 ttttacggca gctgggatca agttgatatt cttctttgat ggcatggtgg agcaggataa
     1201 gagagatgaa tgggtgaaac gaaggctcaa gaacaacagg gagatatcca ggatttttca
     1261 ttacatcaag tcacacaagg agcagccagg cagaaatatg ttcttcatcc cctcagggct
     1321 agctgtgttt acacgatttg ctctaaagac actgggccag gaaactttgt gttctttgca
     1381 ggaagcagat tatgaggtag cttcctatgg cctccagcat aactgtcttg ggattctggg
     1441 ggaagacact gattacctaa tctatgacac ttgtccctac ttttcaatta gcgagctctg
     1501 cctagagagc ctggacaccg tcatgctctg cagagagaag ctctgtgaga gtctgggcct
     1561 ctgtgtggcc gaccttcctc ttctggcctg cctccttggc aacgacataa tcccagaggg
     1621 catgtttgaa agctttaggt acaaatgctt atcgtcctac acctctgtaa aagagaactt
     1681 tgacaaaaaa ggtaacatca tattagctgt gtcagaccat atatcgaaag ttctttactt
     1741 gtatcaaggt gagaaaaaat tagaagagat attacctctg ggaccaaaca aagctctttt
     1801 ttataaagga atggcatcat atcttttacc aggacaaaaa tctccatggt ttttccaaaa
     1861 acccaaaggt gtaataactt tggacaaaca agtaatatcc acgagttcag acgccgaatc
     1921 cagggaagaa gttcccatgt gttcagatgc tgaatccagg caagaagttc ccatgtgtac
     1981 aggccctgaa tccaggcgag aagttcccgt gtatacaaat tctgaaccca ggcaagaagt
     2041 tcccatgtgt tcagaccctg aacccaggca agaagttccc acgtgtacag gccctgaatc
     2101 caggcgagaa gttcccatgt gttcagaccc tgaacccagg caagaagttc ccatgtgtac
     2161 aggccctgaa gccaggcaag aagttcccat gtatacagac tctgaaccca ggcaagaagt
     2221 tcccatgtat acagactctg aacccaggca agaagttccc atgtatacag gctctgaacc
     2281 caggcaagaa gttcccatgt atacaggccc tgaatccagg caagaagttc ccatgtatac
     2341 aggccctgaa tccaggcaag aagttttaat acggacagac cctgaatcta ggcaagaaat
     2401 tatgtgtaca ggccatgaat ccaaacagga agttcccata tgtacagatc ctatatccaa
     2461 gcaagaagac tccatgtgta cacacgctga aatcaatcaa aaattacctg tagcaacaga
     2521 ttttgaattt aagctagaag ctctcatgtg tacaaaccct gaaattaaac aagaagaccc
     2581 cacaaatgtg gggcctgaag taaagcaaca agtaaccatg gtttcagaca ctgaaatctt
     2641 aaaggttgct agaacacatc acgtccaagc agaaagctac ctggtgtaca acatcatgag
     2701 cagtggagag attgaatgca gcaacaccct agaagatgag cttgaccagg ccttacccag
     2761 ccaggccttc atttaccgtc ccattcgaca gcgggtctac tcactcttac tggaggactg
     2821 tcaagatgtc accagcacct gcctagctgt caaggagtgg tttgtgtatc ctgggaaccc
     2881 actgaggcac ccggacctcg tcaggccgct gcagatgacc attccagggg gaacgcctag
     2941 tttgaaaata ttatggctga accaagagcc agaaatacag gttcggcgct tggacacact
     3001 cctagcctgt ttcaatcttt cctcctcaag agaagagctg caggctgtcg agagcccatt
     3061 tcaagctttg tgctgcctct tgatctacct ctttgtccag gtggacacgc tttgcctgga
     3121 ggatttgcat gcgtttattg cgcaggcctt gtgcctccaa ggaaaatcca cctcgcagct
     3181 tgtaaatcta cagcctgatt acatcaaccc cagagccgtg cagctgggct cccttctcgt
     3241 ccgcggcctc accactctgg ttttagtcaa cagcgcatgt ggcttcccct ggaagacgag
     3301 tgatttcatg ccctggaatg tatttgacgg gaagcttttt catcagaagt acttgcaatc
     3361 tgaaaagggt tatgctgtgg aggttctttt agaacaaaat agatctcggc tcaccaaatt
     3421 ccacaacctg aaggcagtcg tctgcaaggc ctgcatgaag gagaacagac gcatcactgg
     3481 ccgagcccac tggggctcac accacgcagg gaggtgggga agacagggct ccagctacca
     3541 caggacgggc tctgggtata gccgttccag tcagggacag ccgtggagag accagggacc
     3601 aggaagcaga cagtatgagc atgaccagtg gagaaggtac ggatccgact acaaggacga
     3661 cgatgacaaa taggaattca tcgatggccg ccatggccca acttgtttat tgcagcttat
     3721 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3781 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgggaat
     3841 taattcggcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     3901 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4141 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4201 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4261 aagctgttaa cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4321 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4381 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     4441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4501 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4561 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4621 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4681 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4741 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4801 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     4861 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4921 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4981 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5041 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5101 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5161 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5221 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5281 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5341 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5401 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5461 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5521 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5581 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5641 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5701 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5761 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5821 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5881 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5941 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6001 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6061 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6121 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6181 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6241 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6301 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6361 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6421 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6481 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6541 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6601 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6661 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6721 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6781 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6841 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6901 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6961 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7021 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7081 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7141 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7201 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7261 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7321 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac
     7381 gaattaattc gagctcgccc
//
