LOCUS       sequence-305908-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  perform light induced optogenetic clustering of TCR zeta-Chain 
            tagged with yellow fluorescent protein as described in the 
            associated publication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  AUTHORS   Ma Y, Lim YJ, Benda A, Lou J, Goyette J, Gaus K
  TITLE     Clustering of the zeta-Chain Can Initiate T Cell Receptor Signaling.
  JOURNAL   Int J Mol Sci. 2020 May 15;21(10). pii: ijms21103498. doi: 
            10.3390/ijms21103498.
  PUBMED    32429097
REFERENCE   2  (bases 1 to 6843)
  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: "Int J Mol 
            Sci."; date: "2020-05-15"; volume: "21(10). pii: ijms21103498. doi";
            pages: " 10.3390/ijms21103498"
COMMENT     Sequence Label: LIC-Z-YFP
FEATURES             Location/Qualifiers
     source          1..6843
                     /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"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1033..1035,1036..1038,1039..1551)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1078..1099)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1339..1358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1686..1707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1762..3255
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     polyA_signal    3631..3752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3668..3687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3722..3741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3759..4214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3896..3917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4108..4127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4241..4345
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4347..4704
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4347..4538
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4370..4390)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4555..4690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4739..5533
                     /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             4739..5533
                     /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(4793..4812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5403..5422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5721..5740)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5765..5812
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6141..6729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6630..6649
                     /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 gctagcgcta
      601 ccggactcag atctcgaggc caccatggga tgtgtctgca gctcaaaccc tgaaggagga
      661 ggaggggata gagtgaagtt cagcaggagc gcagacgccc ccgcgtacca gcagggccag
      721 aaccagctct ataacgagct caatctagga cgaagagagg agtacgatgt tttggacaag
      781 agacgtggcc gggaccctga gatgggggga aagccgagaa ggaagaaccc tcaggaaggc
      841 ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa
      901 ggcgagcgcc ggaggggcaa ggggcacgat ggcctttacc agggtctcag tacggccacc
      961 aaggacacct acgacgccct tcacatgcag gccctgcccc ctcgcggttc tggaggcggt
     1021 ggaagtggta ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1081 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1141 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     1201 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc
     1261 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1321 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1381 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1441 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1501 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1561 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1621 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     1681 gatcacatgg tcctgctgga gttcgtgacc gccgccggca tggacgagct gtacaagggc
     1741 tcaactggaa gtacaggaac aatgaagatg gacaaaaaga ctatagtttg gtttagaaga
     1801 gacctaagga ttgaggataa tcctgcatta gcagcagctg ctcacgaagg atctgttttt
     1861 cctgtcttca tttggtgtcc tgaagaagaa ggacagtttt atcctggaag agcttcaaga
     1921 tggtggatga aacaatcact tgctcactta tctcaatcct tgaaggctct tggatctgac
     1981 ctcactttaa tcaaaaccca caacacgatt tcagcgatct tggattgtat ccgcgttacc
     2041 ggtgctacaa aagtcgtctt taaccacctc tatgatcctg tttcgttagt tcgggaccat
     2101 accgtaaagg agaagctggt ggaacgtggg atctctgtgc aaagctacaa tggagatcta
     2161 ttgtatgaac cgtgggagat atactgcgaa aagggcaaac cttttacgag tttcaattct
     2221 tactggaaga aatgcttaga tatgtcgatt gaatccgtta tgcttcctcc tccttggcgg
     2281 ttgatgccaa taactgcagc ggctgaagcg atttgggcgt gttcgattga agaactaggg
     2341 ctggagaatg aggccgagaa accgagcaat gcgttgttaa ctagagcttg gtctccagga
     2401 tggagcaatg ctgataagtt actaaatgag ttcatcgaga agcagttgat agattatgca
     2461 aagaacagca agaaagttgt tgggaattct acttcactac tttctccgta tctccatttc
     2521 ggggaaataa gcgtcagaca cgttttccag tgtgcccgga tgaaacaaat tatatgggca
     2581 agagataaga acagtgaagg agaagaaagt gcagatcttt ttcttagggg aatcggttta
     2641 agagagtatt ctcggtatat atgtttcaac ttcccgttta ctcacgagca atcgttgttg
     2701 agtcatcttc ggtttttccc ttgggatgct gatgttgata agttcaaggc ctggagacaa
     2761 ggcaggaccg gttatccgtt ggtggatgcc ggaatgagag agctttgggc taccggatgg
     2821 atgcataaca gaataagagt gattgtttca agctttgctg tgaagtttct tctccttcca
     2881 tggaaatggg gaatgaagta tttctgggat acacttttgg atgctgattt ggaatgtgac
     2941 atccttggct ggcagtatat ctctgggagt atccccgatg gccacgagct tgatcgcttg
     3001 gacaatcccg cgttacaagg cgccaaatat gacccagaag gtgagtacat aaggcaatgg
     3061 cttcccgagc ttgcgagatt gccaactgaa tggatccatc atccatggga cgctccttta
     3121 accgtactca aagcttctgg tgtggaactc ggaacaaact atgcgaaacc cattgtagac
     3181 atcgacacag ctcgtgagct actagctaaa gctatttcaa gaacccgtga agcacagatc
     3241 atgatcggag cagcataagc ggccgcgact ctagatcata atcagccata ccacatttgt
     3301 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3361 gaatgcaatt gttgttggtc aaagcttctg gtgtggaact cggaacaaac tatgcgaaac
     3421 ccattgtaga catcgacaca gctcgtgagc tactagctaa agctatttca agaacccgtg
     3481 aagcacagat catgatcgga gcagcataag cggccgcgac tctagatcat aatcagccat
     3541 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3601 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3661 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3721 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     3781 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3841 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3901 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3961 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4021 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4081 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4141 gctggcaagt atagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4201 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4321 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4381 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4441 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4501 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4561 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4621 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4681 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4741 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4801 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4861 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4921 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4981 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5041 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5101 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5161 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5221 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5281 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5341 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5401 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5461 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5521 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5581 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5641 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5701 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5761 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5821 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5881 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5941 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6001 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6061 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6121 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6181 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6241 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6301 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6361 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6421 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6481 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6541 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6601 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6661 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6721 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6781 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6841 cat
//
