LOCUS       sequence-199319-        7068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Highly efficient anion channelrhodopsin from Rhodomonas sp. 
            (CCAC1630) for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7068)
  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 7068)
  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-R2ACR_142-EYFP
FEATURES             Location/Qualifiers
     source          1..7068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..44)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        29..358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      209..344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     271..290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             425..1219
                     /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(479..498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1089..1108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1395..1516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1432..1451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1486..1505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1565..1581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1565..1581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1578..1600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1589..1605
                     /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(1613..1619,1620..1637,1638..1643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1658..1679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1796..1813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1967..2555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2726..3517,3518..3586))
                     /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     3349..3368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3587..3691)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3766..3785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3957..4336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4337..4540
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4491..4511
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4585..4604
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4585..4603
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5542..5544,5545..6258)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5584..5605)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5845..5864)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6192..6213
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6288..6329
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6339..6356
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6379..6396)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6385..6609
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6655..7068,1..15)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6742..6761)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6952..6973
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
       61 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      181 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      361 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
      421 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      481 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      541 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      601 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      661 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      721 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      781 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      841 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
      901 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
      961 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1021 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1081 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1141 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1201 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     1261 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     1321 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     1381 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1441 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1501 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     1561 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1621 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1681 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1741 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1801 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1861 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2401 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2761 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2821 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2881 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2941 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3001 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3061 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3121 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3181 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3241 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3301 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3361 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3421 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3481 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3541 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3601 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3661 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3721 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     3781 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     3841 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     3901 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     3961 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4021 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4081 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4141 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4201 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4261 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4321 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4381 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4441 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4501 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     4561 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     4621 gttaagcttg gtaccgagct cggatccatg gccaccttta actccacctg gaccacctac
     4681 ggcgcctggg aagctgctaa ccccgagtgc gccgagaaga tgcacaggtc ctccgacgct
     4741 gtgcacgtgg tgcagctcac ctgggccatc gtgagcttcc tgcagagcat gttcatgctg
     4801 tccaggttcc ccaggaacgc ctgggaggcc gtgttcctgc ccctggccga gacatgcctc
     4861 tatggcctgg cctccaccgg caacggctac gtgaccatgg ccaacggcag aacactgccc
     4921 tggggcagga tggccagctg gctggctacc tgccccatta tcatgaacca gatcaacgcc
     4981 gtggccaccg tgaacttcca cggcatcagc ctgaaccgta ttcaagtgtg cgccagcatc
     5041 ctcaccatcg tgatgggcgt gaccgcctcc gtgaccgatg aaccccccat gcagtgggct
     5101 tttttcggcc tgggcaccct gtgcctgatc gtggagtacg tgattaccac catcatcatc
     5161 tccgccggcg tggccaaatt tcagccctac gtggaggaga gcgagacctg taggaaggcc
     5221 atcggcaggc tgtacctggt caggaacatg atcttcagca cctgggccgc cttccctatc
     5281 gtgtggctgc tgtcccccca aggagcctgt gtgttcggcg aggacgtggt gcccctgctg
     5341 tacctgatcg ccgacgccct gtgcaagaac gtgtacggcc tgatctggta taacaccacc
     5401 tgggtgctgc tgggcgatgg agtgtgggac cctaccaagc acgtggccaa gcaagcccct
     5461 gccctgcagc aggagatgac cgccaacacc tccgtgatta tggcccctgc cgtgatccag
     5521 gagaccccct gtgcggccgc cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     5581 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     5641 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     5701 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     5761 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     5821 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     5881 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     5941 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     6001 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     6061 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     6121 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     6181 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     6241 gacgagctgt acaagtaaca tatgctctag agggcccgcg gttcgaaggt aagcctatcc
     6301 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
     6361 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     6421 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     6481 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     6541 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     6601 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     6661 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6721 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6781 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6841 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6901 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6961 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7021 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaa
//
