LOCUS       sequence-464220-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  I-SceI endonuclease expression vector with mammalian promoter to 
            introduce a DSB at a genomic I-SceI site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  AUTHORS   Richardson C, Moynahan ME, Jasin M
  TITLE     Double-strand break repair by interchromosomal recombination: 
            suppression of chromosomal translocations.
  JOURNAL   Genes Dev. 1998 Dec 15. 12(24):3831-42.
  PUBMED    9869637
REFERENCE   2  (bases 1 to 5668)
  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: "Genes Dev. 
            1998 Dec 15. 12(24):3831-42."
COMMENT     Sequence Label: pCBASceI
FEATURES             Location/Qualifiers
     source          1..5668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1771..1791
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1813..1833
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1852..1878
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2686..2705)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2751..2806
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2805..2824)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3167..3183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3167..3183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3180..3202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3191..3207
                     /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)."
     promoter        complement(join(3215..3221,3222..3239,3240..3245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3260..3281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3340..3535
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3386..3521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3541..3675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3578..3597)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3632..3651
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3743..3760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3914..4502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3994..4013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4673..5464,5465..5533))
                     /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     5296..5315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5534..5638)
                     /gene="bla"
                     /label=AmpR promoter
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 gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc cggggggggg gggggggggg gggggggggg
      541 ggggggsgrg gsggrgrggk gsggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgccycg ggccgggggg ggctcggggg rggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc ctcgaaattc
     1741 agatccgccg ccactatggg atcaagatcg ccaaaaaaga agagaaaggt gccgaagaag
     1801 catgcagcac caccaaaaaa aaaacgaaaa gtagaagacc cacgatttat gtacccatac
     1861 gatgttcctg actatgcggg tatgaaaaac atcaaaaaaa accaggtaat gaacctgggt
     1921 ccgaactcta aactgctgaa agaatacaaa tcccagctga tcgaactgaa catcgaacag
     1981 ttcgaagcag gtatcggtct gatcctgggt gatgcttaca tccgttctcg tgatgaaggt
     2041 aaaacctact gtatgcagtt cgagtggaaa aacaaagcat acatggacca cgtatgtctg
     2101 ctgtacgatc agtgggtact gtccccgccg cacaaaaaag aacgtgttaa ccacctgggt
     2161 aacctggtaa tcacctgggg cgcccagact ttcaaacacc aagctttcaa caaactggct
     2221 aacctgttca tcgttaacaa caaaaaaacc atcccgaaca acctggttga aaactacctg
     2281 accccgatgt ctctggcata ctggttcatg gatgatggtg gtaaatggga ttacaacaaa
     2341 aactctacca acaaatcgat cgtactgaac acccagtctt tcactttcga agaagtagaa
     2401 tacctggtta agggtctgcg taacaaattc caactgaact gttacgtaaa aatcaacaaa
     2461 aacaaaccga tcatctacat cgattctatg tcttacctga tcttctacaa cctgatcaaa
     2521 ccgtacctga tcccgcagat gatgtacaaa ctgccgaaca ctatctcctc cgaaactttc
     2581 ctgaaataat aagtcgatcg aggaattcac tcctcaggtg caggctgcct atcagaaggt
     2641 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     2701 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     2761 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2821 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     2881 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     2941 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3001 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     3061 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     3121 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     3181 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3241 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3301 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     3361 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3421 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3481 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3541 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3601 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3661 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4321 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4681 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4741 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4801 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4861 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4921 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4981 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5041 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5101 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5161 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5221 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5281 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5341 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5401 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5461 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5521 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5581 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5641 catttccccg aaaagtgcca cctgggtc
//
