LOCUS       sequence-402935-        6760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SspB-Talin(1974-2541)-mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  AUTHORS   Yu M, Le S, Barnett S, Guo Z, Zhong X, Kanchanawong P, Yan J
  TITLE     Implementing Optogenetic Modulation in Mechanotransduction
  JOURNAL   Phys. Rev. X 10, 021001 – Published 1 April, 2020
REFERENCE   2  (bases 1 to 6760)
  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: "Phys. Rev. 
            X 10, 021001 – Published 1 April, 2020"
COMMENT     Sequence Label: SspB-TalinC11-mEmerald
FEATURES             Location/Qualifiers
     source          1..6760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      592..601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             607..936
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     regulatory      2698..2707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2704..2706,2707..2709,2710..3423)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2749..2770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3010..3029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3357..3378
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3545..3666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3582..3601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3636..3655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3673..4128)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3810..3831)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4022..4041
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4155..4259
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4261..4618
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4261..4452
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4284..4304)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4469..4604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4653..5447
                     /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             4653..5447
                     /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(4707..4726)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5317..5336
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5635..5654)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5679..5726
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6055..6643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6544..6563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agccaccatg
      601 gaattcagct ccccgaaacg ccctaagctg ctgcgtgaat attacgattg gctggttgat
      661 aacagcttta ccccatatct ggtggtggat gccacatacc tgggcgtgaa cgtgcccgtg
      721 gagtatgtga aagacggtca gatcgtgctg aatctgtctg caagtgcgac cggcaacctg
      781 caactgacaa atgattttat ccagttcaac gcccgcttta agggcgtgtc tcgtgaactg
      841 tatatcccga tgggtgccgc tctggccatt tacgctcgcg agaacggcga tggtgtgatg
      901 ttcgaaccag aagaaatcta tgacgagctg aatattggtg gtggttctgg tggtagcact
      961 agtgatatcg gtacccaggc ctgcattaca gcagccagtg ctgtgtctgg tatcattgct
     1021 gacctcgaca ccaccatcat gttcgctact gctggcacac ttaaccgtga gggtgctgaa
     1081 acttttgctg accaccggga gggtatctta aagacagcaa aggttcttgt ggaggacacc
     1141 aaggtcctag tgcagaatgc agctgggagc caggagaagt tggcacaagc cgcccagtcc
     1201 tccgtggcca ccattacccg cctcgctgat gtggtcaagc tcggtgcagc cagcctagga
     1261 gccgaagacc ctgaaactca ggtggtgctg atcaatgcag taaaggacgt agccaaggcc
     1321 ctgggtgacc tcatcagcgc tacgaaggct gcagcgggca aagttgggga tgaccctgca
     1381 gtgtggcagc tcaagaactc tgccaaggtg atggtgacca atgtgacatc attgctcaag
     1441 acagtgaagg ctgtggaaga tgaggccacc aaaggcacac gggccctaga ggcaaccaca
     1501 gagcacatac gtcaggaact ggcggtcttc tgttccccag agccacctgc caagacctct
     1561 acccctgaag acttcatccg aatgaccaag ggtattacta tggcaacagc caaagccgtt
     1621 gctgctggca attcctgtcg acaggaagat gtcattgcca cagccaatct gagccgacgg
     1681 gctattgcgg acatgcttcg ggcttgcaag gaagcagctt tccacccaga agtggcgcct
     1741 gatgtacggc tccgagccct gcactatggc cgggagtgtg ccaatggtta cctggaactg
     1801 ctggaccacg tgctgctgac ccttcagaag ccaaacccag acctgaagca gcagctaact
     1861 ggacactcaa agcgggttgc tggctctgtg actgagctca tccaggctgc tgaagccatg
     1921 aaaggaacag agtgggtgga cccagaggac cctactgtca ttgctgagaa tgagctcttg
     1981 ggagccgcag ccgccatcga ggctgcagcc aagaagctgg agcagctgaa gcccagggcc
     2041 aaacccaagg aggcggatga gtccttgaac tttgaggaac aaatcctaga agctgccaag
     2101 tccatcgctg cagccaccag tgcactggta aaggctgcgt cagcagccca gagggagctg
     2161 gtggctcaag gaaaggtggg cgccattcca gccaatgcac tggatgatgg gcagtggtcg
     2221 cagggcctca tttccgctgc ccgtatggtg gctgcggcca ccaacaatct gtgtgaagca
     2281 gctaacgcag ctgtccaggg ccatgctagc caggagaaac tcatttcctc agccaagcaa
     2341 gtagctgcct ccacagctca gctcctggta gcttgcaagg tcaaagcaga tcaggactct
     2401 gaggcaatga aacggctcca ggctgctggc aatgcagtga agagggcctc agataacctg
     2461 gtaaaggcgg cccagaaagc tgcagccttc gaagaccagg agaatgagac ggtggtggtg
     2521 aaggagaaaa tggttggggg cattgcccag attatcgcag cacaggaaga gatgcttcgg
     2581 aaggaacgag agctggaaga ggctcggaaa aagctcgccc agatccggca gcagcagtac
     2641 aagttcttgc cttcagagct tcgagacgag cacggctccg cgtcccgatc accggtcgcc
     2701 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2761 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2821 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2881 accctcgtga ccaccttgac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     2941 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3001 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3061 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3121 cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag
     3181 aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc
     3241 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3301 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     3361 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3421 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3481 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3541 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3601 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3661 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3721 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3781 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3841 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3901 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3961 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4021 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4081 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4141 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4201 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4261 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4321 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4381 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4441 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4501 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4561 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4621 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4681 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4741 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4801 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4861 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4921 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4981 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5041 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5101 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5161 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5221 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5281 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5341 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5401 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5461 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5521 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5581 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5641 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5701 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5761 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5821 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5881 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5941 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6001 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6061 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6181 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6721 ccctgattct gtggataacc gtattaccgc catgcattag
//
