LOCUS       sequence-435311-        6885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of mNeonGreen, SSLuc, and ChRmine for bioluminescent 
            optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Slaviero AN, Gorantla N, Simkins J, Crespo EL, Ikefuama EC, Tree MO,
            Prakash M, Bjorefeldt A, Barnett LM, Lambert GG, Lipscombe D, Moore 
            CI, Shaner NC, Hochgeschwender U
  TITLE     Engineering luminopsins with improved coupling efficiencies.
  JOURNAL   Neurophotonics. 2024 Apr;11(2):024208. doi: 
            10.1117/1.NPh.11.2.024208. Epub 2024 Mar 29.
  PUBMED    38559366
REFERENCE   2  (bases 1 to 6885)
  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: 
            "Neurophotonics."; date: "2024-04"; volume: "11(2):024208. doi"; 
            pages: " 10.1117/1.NPh.11.2.024208. Epub 2024 Mar 29"
COMMENT     Sequence Label: pAAV-hSyn-LMO11 (GeNL_SS-ChRmine)
FEATURES             Location/Qualifiers
     source          1..6885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   44..173
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        218..665
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      685..694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      994..1003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3007..3024
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3055..3643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3108..3128)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3526..3537)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3675..4151
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3874..3893)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4191..4331
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4191..4320
                     /label=AAV2 ITR
     rep_origin      4406..4861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4493..4512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4703..4724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4878..4897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4997..5019
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5057..5075)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5143..5247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5248..5316,5317..6108)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5466..5485)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6279..6867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6768..6787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      121 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
      181 cacgcgtgtg tctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      241 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      301 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      361 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      421 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      481 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      541 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      601 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      661 cgcagtcgag aaggtaccgg atccgccacc atgggagtca aagttctgtt tgccctgatc
      721 tgcatcgctg tggccgaggc cgtgagcaag ggcgaggagg ataacatggc tagcctccca
      781 gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt
      841 cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt
      901 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
      961 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggatac
     1021 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     1081 acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct
     1141 gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact
     1201 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc
     1261 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     1321 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc
     1381 gagctcaact tcaaggagtg gcaaaaggcc tttaccggat ttgaagactt cgttggtgac
     1441 tgggaacaga tcgctgctta caacctggac caggttctgg aacagggtgg tgtttcttct
     1501 gttctgcaga ccctggctgt ttctgttacc cctatccagc gtatcgttcg ttctggtgaa
     1561 aacggtctga agatcgacat ccatgttatc atcccttacg aaggtctgtc tgctgaccag
     1621 atggctcata tcgaagaagt tttcaaggtt gtttaccctg ttgacgacca tcatttcaag
     1681 gttatcatgg aatacggtac cctggttatc gacggtgtta cccctaacat gctgaactac
     1741 ttcggtcgtc cttacgaagg tatcgctgtt ttcgacggta ataagatcac cgttaccggt
     1801 accctgtgga acggtaacaa gatcatcgac gaacgtctga tcacccctga aggttctatg
     1861 ctgttccgtg ttaccatcaa cggtgttacc ggttaccgtc tgctgaagaa gatctctaac
     1921 ctcgagggcg acccactagt ccagtgtggt ggaattgccg gatccgccac cgcacacgca
     1981 ccaggcaccg accagatgtt ctacgtgggc acaatggacg gctggtatct ggataccaag
     2041 ctgaactccg tggccatcgg cgcccactgg tcttgcttta tcgtgctgac aatcaccaca
     2101 ttctacctgg gctatgagtc ctggacatct cggggcccta gcaagagaac ctccttttac
     2161 gccggctatc aggaggagca gaacctggcc ctgtttgtga atttctttgc catgctgagc
     2221 tacttcggca agatcgtggc cgacacactg ggccacaact tcggcgatgt gggccccttc
     2281 atcatcggct tcggcaatta caggtatgcc gactacatgc tgacctgccc aatgctggtg
     2341 tacgatctgc tgtatcagct gagggccccc tatcgcgtgt cttgtagcgc catcatcttt
     2401 gccatcctga tgtctggcgt gctggccgag ttctacgcag agggcgaccc taggctgagg
     2461 aatggcgcct acgcctggta tggctttggc tgtttctggt ttatcttcgc ctactctatc
     2521 gtgatgagca tcgtggccaa gcagtatagc cggctggccc agctggccca ggatacaggc
     2581 gccgagcaca gcctgcacgt gctgaagttc gccgtgttta ccttctccat gctgtggatt
     2641 ctgtttcccc tggtgtgggc catctgccct agaggcttcg gctggatcga cgataactgg
     2701 acagaggtgg cccactgcgt gtgcgacatc gtggccaagt cctgttacgg ctttgccctg
     2761 gcccggttca gaaagaccta tgatgaggag ctgtttcggc tgctggagca gctgggacac
     2821 gacgaggatg agttccagaa gctggagctg gatatgaggc tgagcagcaa tggcgagcgc
     2881 ctgcggagac tgtctgcggc cgccaagagc aggatcacca gcgagggcga gtacatcccc
     2941 ctggaccaga tcgacatcaa cgtgtgaacg cgtggcgcgc caatattatg ctagccaatt
     3001 gccggtcatc atcaccatca ccattgagtt taattcgata tcaagcttat cgataatcaa
     3061 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3121 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3181 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3241 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3301 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3361 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3421 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     3481 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3541 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3601 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     3661 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     3721 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     3781 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     3841 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     3901 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     3961 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     4021 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     4081 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     4141 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     4201 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4261 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4321 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4381 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4441 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4501 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4561 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4621 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4681 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     4741 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4801 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4861 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4921 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4981 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5041 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5101 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5161 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5221 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5281 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5341 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5401 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5461 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5521 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5581 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5641 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5701 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5761 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5821 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5881 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5941 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6001 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6061 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6121 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggcc
//
