LOCUS       sequence-158520-        6179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent signal joint substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  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 6179)
  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: 289 RFP/CFP
FEATURES             Location/Qualifiers
     source          1..6179
                     /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             671..1348
                     /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"
     CDS             1359..2036
                     /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"
     regulatory      2119..2128
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2125..2127,2128..2130,2131..2844)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2170..2191)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2431..2450)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2778..2799
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2967..3088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3004..3023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3058..3077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3095..3550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3232..3253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3444..3463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3577..3681
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3683..4040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3683..3874
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3706..3726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3891..4026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4075..4869
                     /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             4075..4869
                     /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(4129..4148)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4739..4758
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5057..5076)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5101..5148
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5477..6065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5966..5985
                     /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 gctagccaag
      601 cttgggctgc aggtcgactg cagggttttt gttccagtct gtagcactgt ggtcgactct
      661 catcgatacc atggcctcct ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg
      721 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      781 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
      841 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc
      901 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      961 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggaag
     1021 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca
     1081 gaagaagact atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgcgct
     1141 gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     1201 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggactc
     1261 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcgc
     1321 cgagggccgc caccacctgt tcctgtagca tcgataccat ggcctcctcc gaggacgtca
     1381 tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg
     1441 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1501 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg
     1561 gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc
     1621 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1681 cccaggactc ctccctgcag gacggaagct tcatctacaa ggtgaagttc atcggcgtga
     1741 acttcccctc cgacggcccc gtaatgcaga agaagactat gggctgggag gcctccaccg
     1801 agcgcctgta cccccgcgac ggcgcgctga agggcgagat ccacaaggcc ctgaagctga
     1861 aggacggcgg ccactacctg gtggagttca agtccatcta catggccaag aagcccgtgc
     1921 agctgcccgg ctactactac gtggactcca agctggacat cacctcccac aacgaggact
     1981 acaccatcgt ggagcagtac gagcgcgccg agggccgcca ccacctgttc ctgtagcatc
     2041 gatgagagga tcccacagtg gtagtactcc actgtctggc tgtacaaaaa ccctcgggat
     2101 ccccggggat caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2161 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2221 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2281 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
     2341 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2401 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2461 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2521 gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat
     2581 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     2641 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2701 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     2761 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2821 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     2881 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2941 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3061 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3121 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3181 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3241 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3301 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3361 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3421 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3481 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3541 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3661 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3961 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4021 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5041 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5101 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5161 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5221 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5281 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5341 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5401 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
