LOCUS       sequence-170200-        5461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent Isce joint substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5461)
  AUTHORS   Neal JA, Xu Y, Abe M, Hendrickson E, Meek K
  TITLE     Restoration of ATM Expression in DNA-PKcs-Deficient Cells Inhibits 
            Signal End Joining.
  JOURNAL   J Immunol. 2016 Apr 1;196(7):3032-42. doi: 10.4049/jimmunol.1501654.
            Epub 2016 Feb 26.
  PUBMED    26921311
REFERENCE   2  (bases 1 to 5461)
  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"; doi: 
            "10.4049/jimmunol.1501654"; journalName: "J Immunol"; date: 
            "2016-04-1- 1"; volume: "196"; issue: "7"; pages: "3032-42"
COMMENT     Sequence Label: isce rfp/cfp
FEATURES             Location/Qualifiers
     source          1..5461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             667..1344
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGALKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(797..815)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(847..867)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1065..1084
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1259..1282
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     regulatory      1401..1410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1407..1409,1410..1412,1413..2126)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1452..1473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1713..1732)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2060..2081
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2249..2370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2286..2305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2340..2359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2377..2832)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2514..2535)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2726..2745
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2859..2963
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2965..3322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2965..3156
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2988..3008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3173..3308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3357..4151
                     /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             3357..4151
                     /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(3411..3430)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4021..4040
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4339..4358)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4383..4430
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4759..5347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5248..5267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcccaa
      601 gcttgggctg caggtcgaag ttacgctagg gataacaggg taatatagtc gactctcatc
      661 gataccatgg cctcctccga ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg
      721 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      781 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      841 gacatcctgt ccccccagtt ccagtacggc tccaaggtgt acgtgaagca ccccgccgac
      901 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      961 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggaagcttc
     1021 atctacaagg tgaagttcat cggcgtgaac ttcccctccg acggccccgt aatgcagaag
     1081 aagactatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgcgctgaag
     1141 ggcgagatcc acaaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1201 tccatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggactccaag
     1261 ctggacatca cctcccacaa cgaggactac accatcgtgg agcagtacga gcgcgccgag
     1321 ggccgccacc acctgttcct gtagcatcga tgagaggatc ctatattacc ctgttatccc
     1381 tagcgtaact gataccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1441 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1501 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1561 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct
     1621 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1681 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1741 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1801 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc cacaacgtct
     1861 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1921 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1981 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2041 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2101 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     2161 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2221 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2341 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     2401 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2461 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2521 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2581 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2641 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2701 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2761 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2821 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2881 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2941 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3001 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3061 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3121 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3241 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3301 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3361 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3421 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3481 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3541 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3601 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3661 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3721 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3781 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3841 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3901 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3961 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4021 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4081 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4141 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4201 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4321 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4381 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4441 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4501 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4561 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     4621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4861 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5341 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5401 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5461 t
//
