LOCUS       sequence-376893-        6678 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 6678)
  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 6678)
  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+IL3 mGluR6)
FEATURES             Location/Qualifiers
     source          1..6678
                     /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    2037..2610
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2060..2612
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2204..2221)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2431..2450
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2630..2639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2636..3343
                     /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    3466..3587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3557..3576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3594..4049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3731..3752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3943..3962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4076..4180
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4182..4539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4182..4373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4205..4225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4390..4525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4574..5368
                     /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             4574..5368
                     /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(4628..4647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5238..5257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5556..5575)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5600..5647
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5976..6564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6465..6484
                     /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 gggcgtgccc gagaccttca acgaggccaa gggcaatggc gagtccctgt ggcagcggca
     1501 gcggctgcag agcgagtgca agatggccaa gatcatgctg ctggtcatcc tcctcttcgt
     1561 gctctcctgg gctccctatt ccgctgtggc cctggtggcc tttgctgggt acgcacacgt
     1621 cctgacaccc tacatgagct cggtgccagc cgtcatcgcc aaggcctctg caatccacaa
     1681 ccccatcatt tacgccatca cccaccccaa gtacagggtg gccattgccc agcacctgcc
     1741 ctgcctgggg gtgctgctgg gtgtatcacg ccggcacagt cgcccctacc ccagctaccg
     1801 ctccacccac cgctccacgc tgaccagcca cacctccaac ctccatccag agcagaatgt
     1861 gcagaagcga aagcggagcc tcaaggccac ctccacggtg gcagccccac ccaagggcga
     1921 ggatgcagag gcccacaaga agagcaggat caccagcgag ggcgagtaca tccccctgga
     1981 ccagatcgac atcaacgtga cagagaccag ccaagtggcg cctgcctaag gatccgcccc
     2041 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2101 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2161 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2221 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     2281 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2341 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2401 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2461 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2521 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2581 gtggttttcc tttgaaaaac acgatgataa tatggccaca ccaccggtcg ccaccatggt
     2641 gggtgaggat agcgtgctga tcaccgagaa catgcacatg aaactgtaca tggagggcac
     2701 cgtgaacgac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     2761 ccagaccatg aagatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     2821 ggctaccagc ttcatgtacg gcagcaaaac ctttatcaac cacacccagg gcatccccga
     2881 cttctttaag cagtccttcc ctgagggctt cacatgggag aggatcacca catacgaaga
     2941 cgggggcgtg ctgaccgcta cccaggacac cagcctccag aacggctgcc tcatctacaa
     3001 cgtcaagatc aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacact
     3061 cggctgggag gccagcaccg agatgctgta ccccgctgac agcggcctga gaggccatag
     3121 ccagatggcc ctgaagctcg tgggcggggg ctacctgcac tgctccctca agaccacata
     3181 cagatccaag aaacccgcta agaacctcaa gatgcccggc ttctacttcg tggacaggag
     3241 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg agatggctgt
     3301 ggccaggtac tgcgacctgc ctagcaaact ggggcacagc tgatgcggcc gcgactctag
     3361 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3421 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3601 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3661 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3721 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3781 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3841 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3901 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3961 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4021 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4081 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4141 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4441 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4561 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4621 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4681 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4741 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4801 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4861 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4921 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4981 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5041 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5101 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5161 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5221 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5281 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5341 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5401 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5461 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5521 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5581 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5641 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5701 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5761 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6661 cgtattaccg ccatgcat
//
