LOCUS       sequence-376892-        6582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic tool.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  AUTHORS   Kralik J, van Wyk M, Stocker N, Kleinlogel S
  TITLE     Bipolar cell targeted optogenetic gene therapy restores parallel 
            retinal signaling and high-level vision in the degenerated retina.
  JOURNAL   Commun Biol. 2022 Oct 20;5(1):1116. doi: 10.1038/s42003-022-04016-1.
  PUBMED    36266533
REFERENCE   2  (bases 1 to 6582)
  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: "Commun 
            Biol."; date: "2022-10-20"; volume: "5(1):1116. doi"; pages: " 
            10.1038/s42003-022-04016-1"
COMMENT     Sequence Label: pIRES2_hMela(CT mGluR6)
FEATURES             Location/Qualifiers
     source          1..6582
                     /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"
     misc_feature    1941..2514
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1964..2516
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2108..2125)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2335..2354
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2534..2543
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2540..3247
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     polyA_signal    3370..3491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3461..3480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3498..3953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3635..3656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3847..3866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4086..4443
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4086..4277
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4109..4129)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4294..4429
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4478..5272
                     /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             4478..5272
                     /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(4532..4551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5142..5161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5460..5479)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5504..5551
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5880..6468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6369..6388
                     /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 atctcgagct caagcttcga attcgccacc atgaaccctc cttcggggcc
      661 aagagtcccg cccagcccaa cccaagagcc cagctgcatg gccaccccag caccacccag
      721 ctggtgggac agctcccaga gcagcatctc cagcctgggc cggcttccat ccatcagtcc
      781 cacagcacct gggacttggg ctgctgcctg ggtccccctc cccacggttg atgttccaga
      841 ccatgcccac tataccctgg gcacagtgat cttgctggtg ggactcacgg ggatgctggg
      901 caacctgacg gtcatctata ccttctgcag gagcagaagc ctccggacac ctgccaacat
      961 gttcattatc aacctcgcgg tcagcgactt cctcatgtcc ttcacccagg cccctgtctt
     1021 cttcaccagt agcctctata agcagtggct ctttggggag acaggctgcg agttctatgc
     1081 cttctgtgga gctctctttg gcatttcctc catgatcacc ctgacggcca tcgccctgga
     1141 ccgctacctg gtaatcacac gcccgctggc cacctttggt gtggcgtcca agaggcgtgc
     1201 ggcatttgtc ctgctgggcg tttggctcta tgccctggcc tggagtctgc cacccttctt
     1261 cggctggagc gcctacgtgc ccgaggggtt gctgacatcc tgctcctggg actacatgag
     1321 cttcacgccg gccgtgcgtg cctacaccat gcttctctgc tgcttcgtgt tcttcctccc
     1381 tctgcttatc atcatctact gctacatctt catcttcagg gccatccggg agacaggacg
     1441 ggctctccag accttcgggg cctgcaaggg caatggcgag tccctgtggc agcggcagcg
     1501 gctgcagagc gagtgcaaga tggccaagat catgctgctg gtcatcctcc tcttcgtgct
     1561 ctcctgggct ccctattccg ctgtggccct ggtggccttt gctgggtacg cacacgtcct
     1621 gacaccctac atgagctcgg tgccagccgt catcgccaag gcctctgcaa tccacaaccc
     1681 catcatttac gccatcaccc accccaagta cagggtggcc attgcccagc acctgccctg
     1741 cctgttccat ccagagcaga atgtgcagaa gcgaaagcgg agcctcaagg ccacctccac
     1801 ggtggcagcc ccacccaagg gcgaggatgc agaggcccac aagaagagca ggatcaccag
     1861 cgagggcgag tacatccccc tggaccagat cgacatcaac gtgacagaga ccagccaagt
     1921 ggcgcctgcc taaggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     1981 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     2041 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     2101 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     2161 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     2221 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     2281 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     2341 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     2401 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     2461 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     2521 cacaccaccg gtcgccacca tggtgggtga ggatagcgtg ctgatcaccg agaacatgca
     2581 catgaaactg tacatggagg gcaccgtgaa cgaccaccac ttcaagtgca catccgaggg
     2641 cgaaggcaag ccctacgagg gcacccagac catgaagatc aaggtggtcg agggcggccc
     2701 tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca aaacctttat
     2761 caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg
     2821 ggagaggatc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     2881 ccagaacggc tgcctcatct acaacgtcaa gatcaacggg gtgaacttcc catccaacgg
     2941 ccctgtgatg cagaagaaaa cactcggctg ggaggccagc accgagatgc tgtaccccgc
     3001 tgacagcggc ctgagaggcc atagccagat ggccctgaag ctcgtgggcg ggggctacct
     3061 gcactgctcc ctcaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc
     3121 cggcttctac ttcgtggaca ggagactgga aagaatcaag gaggccgaca aagagaccta
     3181 cgtcgagcag cacgagatgg ctgtggccag gtactgcgac ctgcctagca aactggggca
     3241 cagctgatgc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     3301 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3361 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3481 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3541 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3601 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3661 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3721 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3781 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3841 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3901 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4081 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4321 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4381 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4441 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4501 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4561 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4621 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4681 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4741 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4801 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4861 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4921 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4981 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5041 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5101 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5161 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5221 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5281 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5341 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5401 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5461 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5521 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5581 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5641 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5701 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6541 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
