LOCUS       Exported                5665 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  I-SceI endonuclease expression vector with mammalian promoter to 
            introduce a DSB at a genomic I-SceI site.
ACCESSION   .
VERSION     .
KEYWORDS    pCBASceI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5665)
  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 5665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5665
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1772..1792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1814..1834
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1853..1879
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2687..2706)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2752..2807
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2806..2825)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3168..3184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3168..3184)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3181..3203)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3192..3208
                     /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(3216..3246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3261..3282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3341..3536
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3387..3522
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3449..3468
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3542..3676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3579..3598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3633..3652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3744..3761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3915..4503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3995..4014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4674..5534)
                     /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     5297..5316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5535..5639)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg ggggggsgcg ccrggsgggg gggggggggg gggggggggg
      541 gggggggsgr ggsggrgrgg kgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgccyc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgaaatt
     1741 cagatccgcc gccactatgg gatcaagatc gccaaaaaag aagagaaagg tgccgaagaa
     1801 gcatgcagca ccaccaaaaa aaaaacgaaa agtagaagac ccacgattta tgtacccata
     1861 cgatgttcct gactatgcgg gtatgaaaaa catcaaaaaa aaccaggtaa tgaacctggg
     1921 tccgaactct aaactgctga aagaatacaa atcccagctg atcgaactga acatcgaaca
     1981 gttcgaagca ggtatcggtc tgatcctggg tgatgcttac atccgttctc gtgatgaagg
     2041 taaaacctac tgtatgcagt tcgagtggaa aaacaaagca tacatggacc acgtatgtct
     2101 gctgtacgat cagtgggtac tgtccccgcc gcacaaaaaa gaacgtgtta accacctggg
     2161 taacctggta atcacctggg gcgcccagac tttcaaacac caagctttca acaaactggc
     2221 taacctgttc atcgttaaca acaaaaaaac catcccgaac aacctggttg aaaactacct
     2281 gaccccgatg tctctggcat actggttcat ggatgatggt ggtaaatggg attacaacaa
     2341 aaactctacc aacaaatcga tcgtactgaa cacccagtct ttcactttcg aagaagtaga
     2401 atacctggtt aagggtctgc gtaacaaatt ccaactgaac tgttacgtaa aaatcaacaa
     2461 aaacaaaccg atcatctaca tcgattctat gtcttacctg atcttctaca acctgatcaa
     2521 accgtacctg atcccgcaga tgatgtacaa actgccgaac actatctcct ccgaaacttt
     2581 cctgaaataa taagtcgatc gaggaattca ctcctcaggt gcaggctgcc tatcagaagg
     2641 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     2701 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     2761 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     2821 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     2881 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     2941 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3001 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3061 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3121 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     3181 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3241 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3301 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     3361 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3421 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3481 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3541 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3661 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3721 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4081 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4201 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4261 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4321 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4381 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4441 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4501 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4561 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4621 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4921 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4981 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5041 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5101 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5161 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5221 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5281 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5341 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5401 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5461 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5521 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5581 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5641 acatttcccc gaaaagtgcc acctg
//
