LOCUS       sequence-384406-        7927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for iLMO2, an opto-chemogenetic probe 
            for neuronal inhibition by light or chemical.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7927)
  AUTHORS   Tung JK, Gutekunst CA, Gross RE
  TITLE     Inhibitory luminopsins: genetically-encoded bioluminescent opsins 
            for versatile, scalable, and hardware-independent optogenetic 
            inhibition.
  JOURNAL   Sci Rep. 2015 Sep 24;5:14366. doi: 10.1038/srep14366.
  PUBMED    26399324
REFERENCE   2  (bases 1 to 7927)
  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: "Sci Rep."; 
            date: "2015-09-24"; volume: "5:14366. doi"; pages: " 
            10.1038/srep14366"
COMMENT     Sequence Label: pcDNA3-iLMO2
FEATURES             Location/Qualifiers
     source          1..7927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1815..1817,1818..2330)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(1857..1878)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2118..2137)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2465..2486
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3480..3498)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3481..3498)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3518..3535)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3524..3748
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3794..4222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3881..3900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4091..4112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4231..4251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4236..4565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4416..4551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4478..4497
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4632..5426
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4686..4705)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5296..5315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5600..5721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5637..5656)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5691..5710
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5770..5786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5770..5786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5783..5805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5794..5810
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5818..5824,5825..5842,5843..5848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5863..5884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6001..6018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6172..6760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6252..6271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6931..7722,7723..7791))
                     /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     7554..7573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7792..7896)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccccgggtac cggtaggcca ccatgacaga gaccctgcct cccgtgaccg
      961 agagtgccgt ggcccttcaa gccgaggtta cccaaaggga gttgttcgag ttcgtgctga
     1021 acgacccttt gcttgcaagc agtctctata tcaacatcgc acttgcagga ctgagtatac
     1081 tgctgttcgt ttttatgacc cgaggactcg atgatccacg ggcaaaactt attgctgtgt
     1141 caaccatcct tgtgcctgtc gtcagcattg cctcctacac tggattggcg agcggcctga
     1201 caatttccgt tcttgaaatg ccagcgggcc attttgcaga aggcagctca gtgatgctgg
     1261 gaggagaaga ggtagatggt gtagtcacca tgtggggacg gtatctcacc tgggcacttt
     1321 ccacgcccat gattctcctc gctctgggtc tcctggccgg aagcaatgct acaaagctct
     1381 tcacagctat cactttcgat atcgctatgt gcgtgactgg ccttgccgcg gccctgacta
     1441 cctcctccca cctcatgaga tggttctggt acgctatcag ttgtgcatgc tttctggtgg
     1501 tcttgtatat cctgctggtg gagtgggcac aggacgccaa agccgcggga accgctgaca
     1561 tgttcaatac cctgaagctg ttgacagtag tgatgtggct ggggtatcca attgtgtggg
     1621 ctcttggagt cgagggtatc gcggtgttgc ccgttggggt gacgagctgg ggatattctt
     1681 tcctggatat cgtggcaaag tacattttcg cattcttgct cctgaactat ctgacgtcaa
     1741 acgaatctgt cgtgtccggc agcattttgg atgttccatc tgcttctggg accccggctg
     1801 atgatgcggc cgcagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1861 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1921 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1981 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
     2041 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2101 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2161 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2221 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     2281 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     2341 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2401 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2461 atcacatggt cctgctggag ttcgtgaccg ccgccggggg taccaaggtg tacgaccccg
     2521 agcagaggaa gaggatgatc accggccccc agtggtgggc caggtgcaag cagatgaacg
     2581 tgctggacag cttcatcaac tactacgaca gcgagaagca cgccgagaac gccgtgatct
     2641 tcctgcacgg caacgccact agcagctacc tgtggaggca cgtggtgccc cacatcgagc
     2701 ccgtggccag gtgcatcatc cccgatctga tcggcatggg caagagcggc aagagcggca
     2761 acggcagcta caggctgctg gaccactaca agtacctgac cgcctggttc gagcttctga
     2821 acctgcccaa gaagatcatc ttcgtgggcc acgactgggg cgccgccctg gccttccact
     2881 acgcctacga gcaccaggac aggatcaagg ccatcgtgca catggagagc gtggtggacg
     2941 tgatcgagag ctgggacgag tggccagaca tcgaggagga catcgccctg atcaagagcg
     3001 aggagggcga gaagatggtg ctggagaaca acttcttcgt ggagaccgtg ctgcccagca
     3061 agatcatgag aaagctggag cccgaggagt tcgccgccta cctggagccc ttcaaggaga
     3121 agggcgaggt gagaagaccc accctgagct ggcccagaga gatccccctg gtgaagggcg
     3181 gcaagcccga cgtggtgcag atcgtgagaa actacaacgc ctacctgaga gccagcgacg
     3241 acctgcccaa gctgttcatc gagggcgacc ccggcttctt cagcaacgcc atcgtggagg
     3301 gcgccaagaa gttccccaac accgagttcg tgaaggtgaa gggcctgcac ttcctccagg
     3361 aggacgcccc cgacgagatg ggcaagtaca tcaagagctt cgtggagaga gtgctgaaga
     3421 acgagcagtt ctgctacgag aacgaggtgt aactcgagca tgcatctaga gggccctatt
     3481 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     3541 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3601 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3661 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3721 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3781 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3841 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3901 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3961 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4021 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4081 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4141 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4201 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4441 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4501 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4561 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4621 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4681 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4741 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4801 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4861 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4921 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4981 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5041 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5101 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5161 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5221 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5281 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5341 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5401 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5461 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5521 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5581 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5641 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5701 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5761 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5821 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5881 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5941 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7921 tgacgtc
//
