LOCUS       sequence-170199-        6837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent coding joint substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6837)
  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 6837)
  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: 290 RFP/CFP
FEATURES             Location/Qualifiers
     source          1..6837
                     /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             654..1331
                     /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             1342..2019
                     /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             2030..2707
                     /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      2777..2786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2783..2785,2786..2788,2789..3502)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2828..2849)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3089..3108)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3436..3457
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3625..3746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3662..3681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3716..3735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3753..4208)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3890..3911)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4102..4121
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4235..4339
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4341..4698
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4341..4532
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4364..4384)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4549..4684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4733..5527
                     /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             4733..5527
                     /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(4787..4806)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5397..5416
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5715..5734)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5759..5806
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6135..6723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6624..6643
                     /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 gctagctcga
      601 ccacagtgct acagactgga acaaaaaccc tgcagtcgac tctcatcgat accatggcct
      661 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
      721 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
      781 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
      841 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
      901 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
      961 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg aagcttcatc tacaaggtga
     1021 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct
     1081 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgc gctgaagggc gagatccaca
     1141 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg
     1201 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct
     1261 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc
     1321 tgttcctgta gcatcgatac catggcctcc tccgaggacg tcatcaagga gttcatgcgc
     1381 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1441 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     1501 ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg
     1561 aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg
     1621 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1681 caggacggaa gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc
     1741 cccgtaatgc agaagaagac tatgggctgg gaggcctcca ccgagcgcct gtacccccgc
     1801 gacggcgcgc tgaagggcga gatccacaag gccctgaagc tgaaggacgg cggccactac
     1861 ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac
     1921 tacgtggact ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag
     1981 tacgagcgcg ccgagggccg ccaccacctg ttcctgtagc atcgatacca tggcctcctc
     2041 cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     2101 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2161 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     2221 gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa
     2281 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2341 ggtgaccgtg acccaggact cctccctgca ggacggaagc ttcatctaca aggtgaagtt
     2401 catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagacta tgggctggga
     2461 ggcctccacc gagcgcctgt acccccgcga cggcgcgctg aagggcgaga tccacaaggc
     2521 cctgaagctg aaggacggcg gccactacct ggtggagttc aagtccatct acatggccaa
     2581 gaagcccgtg cagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca
     2641 caacgaggac tacaccatcg tggagcagta cgagcgcgcc gagggccgcc accacctgtt
     2701 cctgtagcat cgatgagagg atcccgaggg tttttgtaca gccagacagt ggagtactac
     2761 cactgtggga ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2821 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2881 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2941 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag
     3001 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3061 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3121 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3181 ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat
     3241 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     3301 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3361 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3421 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3481 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     3541 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3601 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3661 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3721 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3781 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3841 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3901 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3961 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4021 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4081 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4141 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4201 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4321 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4381 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4441 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4561 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4621 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4681 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4741 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4801 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4861 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4921 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4981 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5041 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5101 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5161 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5221 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5281 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5341 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5401 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5461 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5521 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5581 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5641 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5701 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5761 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5821 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5881 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5941 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6241 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6781 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
