LOCUS       sequence-305901-        6595 bp    DNA     circular SYN 14-APR-2025
DEFINITION  perform light induced optogenetic clustering of TCR zeta-Chain as 
            described in the associated publication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6595)
  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 6595)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..6595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      560..569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             974..1681
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1119..1137)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1387..1406
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1706..3199
                     /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    3324..3445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3361..3380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3415..3434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3452..3907)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3589..3610)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3801..3820
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3934..4038
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4040..4397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4040..4231
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4063..4083)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4248..4383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4432..5226
                     /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             4432..5226
                     /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(4486..4505)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5096..5115
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5414..5433)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5458..5505
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6323..6342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggactca gatctcgagg ccaccatggg atgtgtctgc agctcaaacc ctgaaggagg
      601 aggaggggat agagtgaagt tcagcaggag cgcagacgcc cccgcgtacc agcagggcca
      661 gaaccagctc tataacgagc tcaatctagg acgaagagag gagtacgatg ttttggacaa
      721 gagacgtggc cgggaccctg agatgggggg aaagccgaga aggaagaacc ctcaggaagg
      781 cctgtacaat gaactgcaga aagataagat ggcggaggcc tacagtgaga ttgggatgaa
      841 aggcgagcgc cggaggggca aggggcacga tggcctttac cagggtctca gtacggccac
      901 caaggacacc tacgacgccc ttcacatgca ggccctgccc cctcgcggtt ctggaggcgg
      961 tggaagtggt accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     1021 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1081 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1141 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     1201 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     1261 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1321 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1381 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     1441 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     1501 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     1561 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     1621 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1681 gggctcaact ggaagtacag gaacaatgaa gatggacaaa aagactatag tttggtttag
     1741 aagagaccta aggattgagg ataatcctgc attagcagca gctgctcacg aaggatctgt
     1801 ttttcctgtc ttcatttggt gtcctgaaga agaaggacag ttttatcctg gaagagcttc
     1861 aagatggtgg atgaaacaat cacttgctca cttatctcaa tccttgaagg ctcttggatc
     1921 tgacctcact ttaatcaaaa cccacaacac gatttcagcg atcttggatt gtatccgcgt
     1981 taccggtgct acaaaagtcg tctttaacca cctctatgat cctgtttcgt tagttcggga
     2041 ccataccgta aaggagaagc tggtggaacg tgggatctct gtgcaaagct acaatggaga
     2101 tctattgtat gaaccgtggg agatatactg cgaaaagggc aaacctttta cgagtttcaa
     2161 ttcttactgg aagaaatgct tagatatgtc gattgaatcc gttatgcttc ctcctccttg
     2221 gcggttgatg ccaataactg cagcggctga agcgatttgg gcgtgttcga ttgaagaact
     2281 agggctggag aatgaggccg agaaaccgag caatgcgttg ttaactagag cttggtctcc
     2341 aggatggagc aatgctgata agttactaaa tgagttcatc gagaagcagt tgatagatta
     2401 tgcaaagaac agcaagaaag ttgttgggaa ttctacttca ctactttctc cgtatctcca
     2461 tttcggggaa ataagcgtca gacacgtttt ccagtgtgcc cggatgaaac aaattatatg
     2521 ggcaagagat aagaacagtg aaggagaaga aagtgcagat ctttttctta ggggaatcgg
     2581 tttaagagag tattctcggt atatatgttt caacttcccg tttactcacg agcaatcgtt
     2641 gttgagtcat cttcggtttt tcccttggga tgctgatgtt gataagttca aggcctggag
     2701 acaaggcagg accggttatc cgttggtgga tgccggaatg agagagcttt gggctaccgg
     2761 atggatgcat aacagaataa gagtgattgt ttcaagcttt gctgtgaagt ttcttctcct
     2821 tccatggaaa tggggaatga agtatttctg ggatacactt ttggatgctg atttggaatg
     2881 tgacatcctt ggctggcagt atatctctgg gagtatcccc gatggccacg agcttgatcg
     2941 cttggacaat cccgcgttac aaggcgccaa atatgaccca gaaggtgagt acataaggca
     3001 atggcttccc gagcttgcga gattgccaac tgaatggatc catcatccat gggacgctcc
     3061 tttaaccgta ctcaaagctt ctggtgtgga actcggaaca aactatgcga aacccattgt
     3121 agacatcgac acagctcgtg agctactagc taaagctatt tcaagaaccc gtgaagcaca
     3181 gatcatgatc ggagcagcat aagcggccgc gactctagat cataatcagc cataccacat
     3241 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3301 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3361 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3421 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3481 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3541 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3601 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3661 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3721 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3781 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3841 agtatagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3901 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4021 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4081 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4141 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4201 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4261 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4321 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4381 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4441 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4501 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4561 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4621 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4681 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4741 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4801 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4861 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4921 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4981 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5041 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5101 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5161 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5221 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5281 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5341 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5401 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5461 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5521 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5581 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5641 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     6541 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttcc
//
