LOCUS       sequence-75335-e        6393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SSR1.0 has a one I-SceI target site for generate specific double 
            strand break after I-SceI endonuclease expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6393)
  AUTHORS   Gomez-Cabello D, Jimeno S, Fernandez-Avila MJ, Huertas P
  TITLE     New tools to study DNA double-strand break repair pathway choice.
  JOURNAL   PLoS One. 2013 Oct 14;8(10):e77206. doi: 
            10.1371/journal.pone.0077206.
  PUBMED    24155929
REFERENCE   2  (bases 1 to 6393)
  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: "PLoS One.";
            date: "2013-10-14"; volume: "8(10):e77206. doi"; pages: " 
            10.1371/journal.pone.0077206"
COMMENT     Sequence Label: SeeSaw Reporter (SSR1.0)
FEATURES             Location/Qualifiers
     source          1..6393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1037..1046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1043..1045,1046..1048,1049..1759)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1043..1045,1046..1048,1049..1759)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1088..1109)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1349..1368)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1696..1717
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1834..2422
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1887..1907)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2305..2316)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3181..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3272..3291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3309..3764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3446..3467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3658..3677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3791..3895
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3897..4254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3897..4088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3920..3940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4105..4240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4289..5083
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4289..5083
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4343..4362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4953..4972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5271..5290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5315..5362
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5691..6279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6180..6199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatnnt acgggtcatt agtcatagcc atatatggag ttccgcgtta
       61 catacttacg gtaatggccc gcctggctga ccgcccaacg acccccgccc atgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
      601 caccatggtg agcggcctgc tgaaggagag tatgcgcatc aagatgtaca tggagggcac
      661 cgtgaacggc cactacttca agtgcgaggg cgagggcgac ggcaacccct tcgccggcac
      721 ccagagcatg agaatccacg tgaccgaggg cgcccccctg cccttcgcct tcgacatcct
      781 ggccccctgc tgcgagtacg gcagcaggac cttcgtgcac cacaccgccg agatccccga
      841 cttcttcaag cagagcttcc ccgagggctt cacctgggag agaaccacca cctacgagga
      901 cggcggcatc ctgaccgccc accaggacac cagcctggag ggcaactgcc tgatctacaa
      961 ggtgaaggtg cacggcacca acttccccgc cgacggcccc gtgatgaaga acaagagcgg
     1021 tctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     1081 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1141 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1201 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1261 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1321 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1381 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1441 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1501 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1561 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1621 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1681 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1741 catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga attgtaggga
     1801 taacagggta atcgaattct gcagtcgatc gacaatcaac ctctggatta caaaatttgt
     1861 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1921 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1981 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2041 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2101 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2161 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2221 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     2281 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2341 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2401 tccctttggg ccgcctcccc gcctggaatt cgagctcgtc gactgcagaa ttcgaagctt
     2461 gagctcgaga tctgagtccg ttgttggcct tctcgggcag gtcgctgtac cgggccacgc
     2521 tggcctcgta cagctcgaag tactcgtcct tcttcttccg cagcatctgg agccggatgt
     2581 cggtgaagtg gaagccgggc atggtcaggg cgcgcacggc cttcttgctc cggtagctgg
     2641 tgtagtggtg gcagatcagg tgccggtcgc ccaccttcag ggccatcacg ttccggccgc
     2701 acagcacgcc gttctcgggg tacaccacct cggtgctggg ctcccagccg ccgctcttgt
     2761 tcttcatcac ggggccgtcg gcggggaagt tggtgccgtg caccttcacc ttgtagatca
     2821 ggcagttgcc ctccaggctg gtgtcctggt gggcggtcag gatgccgccg tcctcgtagg
     2881 tggtggttct ctcccaggtg aagccctcgg ggaagctctg cttgaagaag tcggggatct
     2941 cggcggtgtg gtgcacgaag gtcctgctgc cgtactcgca gcagggggcc aggatgtcga
     3001 aggcgaaggg caggggggcg ccctcggtca cgtggattct catgctctgg gtggatccac
     3061 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt acttgcttta
     3121 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3301 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3361 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3421 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3481 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3541 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3601 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3661 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3721 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3781 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3841 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3901 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4441 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4801 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5101 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5221 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5281 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5341 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5401 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5461 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5521 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5821 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6361 gattctgtgg ataaccgtat taccgccatg cat
//
