LOCUS       sequence-220895-        7455 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 7455)
  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 7455)
  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..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3660..3683
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3700..3709
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3713..3847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3750..3769)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3804..3823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3920..4277
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3920..4111
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3943..3963)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4128..4263
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4297..4314)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4297..4313)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4306..4328)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4526..4981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4613..4632)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4823..4844
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4998..5017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5152..5174
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5227..5245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5313..5417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5418..5486,5487..6278)
                     /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(5636..5655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6449..7037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6938..6957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7191..7208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(7361..7366,7367..7384,7385..7391)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7399..7415
                     /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     7404..7426
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7423..7439
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7423..7439
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc taagctttct agaatgggtg tgagaggttt gcaaggattt gtgggaagta
      961 cctgcccaca tatatgtaca gtagtaaatt tcaaagaact ggcagagcac caccgaagca
     1021 agtatcctgg atgtacccct accattgtgg ttgatgccat gtgttgtctc agatattggt
     1081 atactccaga atcttggatc tgcggtggcc agtggcgaga atacttttct gctttgcgag
     1141 attttgttaa aacttttacg gcagctggga tcaagttgat attcttcttt gatggcatgg
     1201 tggagcagga taagagagat gaatgggtga aacgaaggct caagaacaac agggagatat
     1261 ccaggatttt tcattacatc aagtcacaca aggagcagcc aggcagaaat atgttcttca
     1321 tcccctcagg gctagctgtg tttacacgat ttgctctaaa gacactgggc caggaaactt
     1381 tgtgttcttt gcaggaagca gattatgagg tagcttccta tggcctccag cataactgtc
     1441 ttgggattct gggggaagac actgattacc taatctatga cacttgtccc tacttttcaa
     1501 ttagcgagct ctgcctagag agcctggaca ccgtcatgct ctgcagagag aagctctgtg
     1561 agagtctggg cctctgtgtg gccgaccttc ctcttctggc ctgcctcctt ggcaacgaca
     1621 taatcccaga gggcatgttt gaaagcttta ggtacaaatg cttatcgtcc tacacctctg
     1681 taaaagagaa ctttgacaaa aaaggtaaca tcatattagc tgtgtcagac catatatcga
     1741 aagttcttta cttgtatcaa ggtgagaaaa aattagaaga gatattacct ctgggaccaa
     1801 acaaagctct tttttataaa ggaatggcat catatctttt accaggacaa aaatctccat
     1861 ggtttttcca aaaacccaaa ggtgtaataa ctttggacaa acaagtaata tccacgagtt
     1921 cagacgccga atccagggaa gaagttccca tgtgttcaga tgctgaatcc aggcaagaag
     1981 ttcccatgtg tacaggccct gaatccaggc gagaagttcc cgtgtataca gattctgaac
     2041 ccaggcaaga agttcccatg tgttcagacc ctgaacccag gcaagaagtt cccacgtgta
     2101 caggccctga atccaggcga gaagttccca tgtgttcaga ccctgaaccc aggcaagaag
     2161 ttcccatgtg tacaggccct gaagccaggc aagaagttcc catgtataca gactctgaac
     2221 ccaggcaaga agttcccatg tatacagact ctgaacccag gcaagaagtt cccatgtata
     2281 caggctctga acccaggcaa gaagttccca tgtatacagg ccctgaatcc aggcaagaag
     2341 ttcccatgta tacaggccct gaatccaggc aagaagtttt aatacggaca gaccctgaat
     2401 ctaggcaaga aattatgtgt acaggccatg aatccaaaca ggaagttccc atatgtacag
     2461 atcctatatc caagcaagaa gactccatgt gtacacacgc tgaaatcaat caaaaattac
     2521 ctgtagcaac agattttgaa tttaagctag aagctctcat gtgtacaaac cctgaaatta
     2581 aacaagaaga ccccacaaat gtggggcctg aagtaaagca acaagtaacc atggtttcag
     2641 acactgaaat cttaaaggtt gctagaacac atcacgtcca agcagaaagc tacctggtgt
     2701 acaacatcat gagcagtgga gagattgaat gcagcaacac cctagaagat gagcttgacc
     2761 aggccttacc cagccaggcc ttcatttacc gtcccattcg acagcgggtc tactcactct
     2821 tactggagga ctgtcaagat gtcaccagca cctgcctagc tgtcaaggag tggtttgtgt
     2881 atcctgggaa cccactgagg cacccggacc tcgtcaggcc gctgcagatg accattccag
     2941 ggggaacgcc tagtttgaaa atattatggc tgaaccaaga gccagaaata caggttcggc
     3001 gcttggacac actcctagcc tgtttcaatc tttcctcctc aagagaagag ctgcaggctg
     3061 tcgaaagccc atttcaagct ttgtgctgcc tcttgatcta cctctttgtc caggtggaca
     3121 cgctttgcct ggaggatttg catgcgttta ttgcgcaggc cttgtgcctc caaggaaaat
     3181 ccacctcgca gcttgtaaat ctacagcctg attacatcaa ccccagagcc gtgcagctgg
     3241 gctcccttct cgtccgcggc ctcaccactc tggttttagt caacagcgca tgtggcttcc
     3301 cctggaagac gagtgatttc atgccctgga atgtatttga cgggaagctt tttcatcaga
     3361 agtacttgca atctgaaaag ggttatgctg tggaggttct tttagaacaa aatagatctc
     3421 ggctcaccaa attccacaac ctgaaggcag tcgtctgcaa ggcctgcatg aaggagaaca
     3481 gacgcatcac tggccgagcc cactggggct cacaccacgc agggaggtgg ggaagacagg
     3541 gctccagcta ccacaggacg ggctctgggt atagccgttc cagtcaggga cagccgtgga
     3601 gagaccaggg accaggaagc agacagtatg agcatgacca gtggagaagg tacggatccg
     3661 actacaagga cgacgatgac aaataggaat tcatcgatgg ccgccatggc ccaacttgtt
     3721 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3781 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3841 ctggatcgat cgggaattaa ttcggcgcag caccatggcc tgaaataacc tctgaaagag
     3901 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3961 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4021 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4081 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4141 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4201 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4261 gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt tacaacgtcg
     4321 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccccttcgc
     4381 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgtagcct
     4441 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4501 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4561 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4621 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4681 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4741 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4801 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4861 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4921 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4981 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca actccgctat
     5041 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     5101 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5161 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagtat tcttgaagac
     5221 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5281 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5341 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5401 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5461 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5521 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5581 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5641 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5701 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5761 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5821 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5881 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5941 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6001 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6061 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6241 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6841 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7321 caattaatgt gagttacctc actcattagg caccccaggc tttacacttt atgcttccgg
     7381 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7441 atgattacga attaa
//
