LOCUS       sequence-406216-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent fusion protein of mNeonGreen, eKL9h, and Volvox 
            channelrhodopsin-1 (luminopsin LMO7) for bioluminescent 
            optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  AUTHORS   Bjorefeldt A, Murphy J, Crespo EL, Lambert GG, Prakash M, Ikefuama 
            EC, Friedman N, Brown TM, Lipscombe D, Moore CI, Hochgeschwender U, 
            Shaner NC
  TITLE     Efficient opto- and chemogenetic control in a single molecule driven
            by FRET-modified bioluminescence.
  JOURNAL   Neurophotonics. 2024 Apr;11(2):021005. doi: 
            10.1117/1.NPh.11.2.021005. Epub 2024 Mar 6.
  PUBMED    38450294
REFERENCE   2  (bases 1 to 7513)
  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):021005. doi"; 
            pages: " 10.1117/1.NPh.11.2.021005. Epub 2024 Mar 6"
COMMENT     Sequence Label: pAAV-EFIa-DIO-LMO7 (mNeonGreen-eKL9h-VChR1)
FEATURES             Location/Qualifiers
     source          1..7513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2486..2495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3452..3461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3783..3816
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3867..3900
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3925..4513
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3978..3998)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4396..4407)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4547..4658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4776..4916
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4776..4905
                     /label=AAV2 ITR
     rep_origin      4991..5446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5078..5097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5288..5309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5463..5482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5582..5604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5642..5660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5728..5832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5833..5901,5902..6693)
                     /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(6051..6070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6864..7452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7353..7372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgcccaatt ccatatgtta ggcggccgcg tcctcctctt cagccaccag tgtctcaacc
     1621 tccatctctt ggccggcgat cgtaattttc tgcttctttc tgatgtcgcc atacaacagg
     1681 atgtgctcgt ggattttgac gcgcaggtaa ttccccagca caccccacat gttctttgca
     1741 atcaggtcga ggatggagtg cccaatggca gatccataag ggctgatatg accgaatcct
     1801 tccgtgccca gcagaaacag gactggaaac attccccaag ccacaaagaa tgtccaggcc
     1861 atcactcgga ccagttctcg acaaatccct tttggaactg tgtggaatgc ttcgatataa
     1921 actttagcag catgaaaata ggtatacata ccatagctca atgagatgag gaagaacagg
     1981 atcttcgtcc accccgtaca catggcgctg gttgcgcccc acacgatgca tcccacatca
     2041 gaaaccaaca ggcccatagt ccttttgcta taatcgtcct tcaggcctgt gaggttgctc
     2101 agatggatca gaaggaccgg gcaggtgagg agccattcgc cataccgcat ccaaactacg
     2161 ccgttccctg aagacagcca cagtgttgca gggctgtcga actcgtggaa ggcctcgatg
     2221 atgcttttca tcatttctat caaggctacg taaacctcct cccatccgca ggtagctcgc
     2281 cacgcttgat acgcatacca ccccaggcaa gccactgaca aagcaaagac tacccactga
     2341 agagtaattg ctattgtctt ttcaatggaa gtgccgcgac tccgaagcca accttcacag
     2401 taacactgtc ctcttggcat gcagacggtg ccgtttccga ggtcggttgg gtagcgaact
     2461 atgaggcttc tagcaacagg gtaatccatg gtggcggatc cggcaattcc accacactgg
     2521 actagtgggt cgccgtttga aatcttctcg aacagccgcc agccgctcac gccgttgatg
     2581 gtcactctga acagcataga gccatcaggg gtgatcagcc gctcgtcgat gatcttattg
     2641 ccgttccaca gggtgcctgt cacggtgatc ttcttgccat cgaacacggc gatgccctcg
     2701 taaggccggc caaaataatt cagcatgttt ggggtcactc cgtcgatgac cagggtgcca
     2761 tactccatga tcactttgaa gtggtgatcg tccacggggt acaccacctt aaacacctcc
     2821 tcgatgtggg ccatctgatc ggcggacagg ccctcgtatg ggatgatcac gtggatgtcg
     2881 atcttcaggc cattctcgcc agagcgcacg atcctctgga taggggtcac ggacacggcc
     2941 agtgtctgca gcacggagct cacgcctccc tgctccagca cctgatccag gttataggcg
     3001 gcggtctgct cccaatcgcc cacgaagtcc tcaaagcctg tgaaggcctt ctgccactcc
     3061 ttgaagttca gctcggtctt gctgtgcttc agctctgtct tgcggaacac gtacatgggc
     3121 tgattcttca gatagttggc ggccattggc ttggcaaagg tatatgtggt cctggctgta
     3181 gatctgtacc gcttgccgtt gcctgtggta tagctccact tgaaggtaga gatgattgtc
     3241 ttatcattgg ggtaggtctt cttggatctg caccagtcgg cggcggtcag gctgtttgtc
     3301 atcacggggc catcggctgg aaagcctgtg cccttcacct gggcctcgcc cttgatgtgg
     3361 gagccctcgt aggtatatct gtagttcact gtcaggctgg cgccgtcctc gaactgcatg
     3421 gtccggtgca cctggtagcc gctgccatcc accatggcgg cctgaaatgg agacatgccg
     3481 tcagggtagg gcagatactg gtggaagcca tagccgatgt gtggcaccag gatccaagga
     3541 gaaaactgca ggtcgccctt ggtggacttc agattcagct cctcgtagcc atcattaggg
     3601 ttgcctgtgc cctggcccac catgtcaaaa tccaccccgt tgatgctccc aaaaatgtgc
     3661 agctcgtgag tagcgggcag actagccata ttatcctcct ctcctttaga cacggcctcg
     3721 gccacagcga tgcagatcag ggcaaacaga actttgactc ccatggtacc gtcgacgcta
     3781 gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac
     3841 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat
     3901 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3961 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4021 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4081 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4141 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4201 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4261 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4321 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     4381 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4441 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4501 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat cttcgactgt gccttctagt
     4561 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4621 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4681 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4741 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4801 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4861 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4921 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4981 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5041 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     5101 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5161 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     5221 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5281 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     5341 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     5401 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5461 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5521 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5581 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5641 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5701 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5761 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6001 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6061 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6121 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6181 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6241 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6301 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6361 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6421 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6481 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6541 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6601 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6661 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6781 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6841 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6901 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6961 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7021 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7081 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7141 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7201 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7261 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7321 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7381 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7441 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7501 tttgctcaca tgt
//
