LOCUS       sequence-199227-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anion channelrhodopsin from Rhodomonas sp. (CCMP768) for expression 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Govorunova EG, Sineshchekov OA, Rodarte EM, Janz R, Morelle O, 
            Melkonian M, Wong GK, Spudich JL
  TITLE     The Expanding Family of Natural Anion Channelrhodopsins Reveals 
            Large Variations in Kinetics, Conductance, and Spectral Sensitivity.
  JOURNAL   Sci Rep. 2017 Mar 3;7:43358. doi: 10.1038/srep43358.
  PUBMED    28256618
REFERENCE   2  (bases 1 to 7065)
  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: "2017-03-3"; volume: "7:43358. doi"; pages: " 
            10.1038/srep43358"
COMMENT     Sequence Label: pcDNA3.1-R1ACR_367-EYFP
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(113..133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        118..447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      298..433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     360..379
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             514..1308
                     /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(568..587)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1178..1197
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1484..1605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1521..1540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1575..1594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1654..1670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1654..1670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1667..1689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1678..1694
                     /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(1702..1708,1709..1726,1727..1732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1747..1768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1885..1902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2056..2644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2136..2155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2815..3606,3607..3675))
                     /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     3438..3457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3676..3780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3855..3874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4046..4425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4426..4629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4580..4600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4674..4693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4674..4692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5628..5630,5631..6344)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5670..5691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5931..5950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6278..6299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6374..6415
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6425..6442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6465..6482)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6471..6695
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6741..7065,1..104)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6828..6847)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7038..7059
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
       61 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
      121 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      181 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
      241 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      301 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      361 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
      421 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
      481 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      541 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      601 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      661 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
      721 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      781 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      841 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      901 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      961 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1021 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     1081 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1141 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1201 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1261 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     1321 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     1381 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1441 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     1501 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1561 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1621 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1681 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2461 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3661 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3781 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     3841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     3901 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     3961 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     4021 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     4081 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     4141 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     4201 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     4261 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     4321 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     4381 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     4441 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4501 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     4561 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4621 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
     4681 actcactata gggagaccca agctggctag ttaagcttgg taccgagctc ggatccatga
     4741 gctacgtgga ggaccctgcc ttcggcttct acggctcctg ggaggccgcc aaccaggact
     4801 gtattgaaag gggcctcagc accaccaaca ataccaggat gatccagctg ctgttcgcct
     4861 ccatctgcgc ctgccaggtc ctgtttatgg ccagcagggc ccctagggtg acatgggagg
     4921 ccgtgtacct gcccgtggtg gagaccgtgc tgtacggcat ggccagcacc ggcaacggca
     4981 gcatgagact ggccgacggc aggatcgtgc ctatcgccag gttctgtagc tggctgatca
     5041 cctgccccat tatgctgttc caggtcgtgg gcatccacga gctgaagtgg aggaccatca
     5101 gctgcaagaa catggtgatg gccgccagcc tgatcagaac cgtgtttggc atcggcgcct
     5161 ccgtgacaat tacccccacc atgagatggg tgcagtactt cctgtccctg tccttcttcc
     5221 tgttcgagct gatcgtggtc taccagatct tcagcgccgg cctgaagaaa ttcggcgccg
     5281 tgaagacaag cctgaacaac atcgtctaca gcaggatcat gttcctgagg ttcctgttct
     5341 tcctgagctg gtccagcttc cccatcatct ggctgctgtc cagcaccggc ttttgtctgg
     5401 tgaacgagga cgtgagcgtg ctggcctacc tggtcgctga catgatctgc aagaacctgt
     5461 acggcatcac caatgccaac accctgttca gggtgctcaa cggcaagtgg acccctgaga
     5521 cagaggagga ggccaagcag cctaaggaca acgagcacgc ccatgtgggc gagacctatt
     5581 tcgagcagat gctgagcggc ctgagaggaa gcaggcacgc ggccgccgtg agcaagggcg
     5641 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     5701 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     5761 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     5821 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     5881 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     5941 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     6001 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     6061 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     6121 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     6181 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     6241 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     6301 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaacatatg ctctagaggg
     6361 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     6421 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     6481 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     6541 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6601 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     6661 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     6721 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6781 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6841 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6901 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6961 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7021 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaac
//
