LOCUS       sequence-467519-        6978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FtACR_mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  AUTHORS   Govorunova EG, Sineshchekov OA, Li H, Gou Y, Chen H, Yang S, Wang Y,
            Mitchell S, Palmateer A, Brown LS, St-Pierre F, Xue MG, Spudich JL
  TITLE     Blue-shifted ancyromonad channelrhodopsins for multiplex 
            optogenetics.
  JOURNAL   Elife. 2025 Sep 19;14:RP106508. doi: 10.7554/eLife.106508.
  PUBMED    40970384
REFERENCE   2  (bases 1 to 6978)
  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: "Elife."; 
            date: "2025-09-19"; volume: "14:RP106508. doi"; pages: " 
            10.7554/eLife.106508"
COMMENT     Sequence Label: FtACR_mCherry_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..6978
                     /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             1739..2446
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1881..1899)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2149..2168
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2476..2517
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2527..2544
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2567..2584)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2573..2797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2843..3271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2930..2949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3140..3161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3280..3300)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3285..3614
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3465..3600
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3527..3546
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3681..4475
                     /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(3735..3754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4345..4364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4651..4772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4688..4707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4742..4761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4821..4837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4821..4837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4834..4856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4845..4861
                     /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(4869..4875,4876..4893,4894..4899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4914..4935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5052..5069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5223..5811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5303..5322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5982..6773,6774..6842))
                     /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     6605..6624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6843..6947)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccatggt gtccggcgag gataatgtgt ctatcgccta
      961 cgccatgatg gtcctggtgt ctggcatcag cgtggtccag gccatctggt acaccatcca
     1021 aacagccaga ggcaagaacc acagctggcg gccagctctg gtcatcggcg ctgaggcctt
     1081 tcagtacatg gtgcggctgg ccgtgtccct gaccgacagc aacatctcca gcggactgac
     1141 cgtgatgaac agcagaggcc tggaagtgga tatcggcacc tacctgggct ggttcttcac
     1201 ctgtcctctg ctggtggacg ccattggcaa tgtggaggac gacatcatct acagcgttga
     1261 gctgtaccgg gccgccatga ccaacaccga ggccgtgtac agccctagaa gagtgtggtg
     1321 gatgagcgcc gtgatgtctg gcatgctgtg tacagtggtg gtggcctctc tggctgaaaa
     1381 gaccacccag atcgccgctg tcttttgcgt gggcggagcc ctgtgcatgg tgctcttcgc
     1441 cggcatcgga agagccatta gacggtgcct cgaccacggc gctgatagac tgacagcata
     1501 tagcctgctg atcgtgttct acacaggctg gagcctgttc cccctgatct gggtgctttc
     1561 tcctctggga tacgacgtgc tggatgccgt gaccgccagg gccctgttcg gcttcgccga
     1621 cctgatcgcc aaaaacctgc tgagcgtgct gctgacaaga ctgaccgtga tgagcgtgaa
     1681 cgccaagcag gctgtgctgg aactgaagat ccagacaagc agccacagcg cggccgccgt
     1741 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     1801 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1861 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1921 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     1981 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2041 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     2101 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     2161 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     2221 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     2281 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     2341 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     2401 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaacata tgctctagag
     2461 ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     2521 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     2581 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2641 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2701 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2761 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2821 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2881 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2941 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3001 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3061 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3121 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3181 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3241 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3361 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3421 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3481 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3541 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3601 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3661 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3721 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3781 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3841 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3901 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3961 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4021 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4081 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4141 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4201 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4261 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4321 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4381 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4441 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4501 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4561 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4621 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4681 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4741 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4801 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4861 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4921 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4981 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5041 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5101 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5161 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5221 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5281 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5341 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5401 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5461 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5521 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5581 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5641 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5701 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5761 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5821 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5881 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5941 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6001 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6061 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6121 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6181 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6241 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6301 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6361 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6421 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6481 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6541 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6601 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6661 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6721 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6781 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6841 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6901 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6961 aaagtgccac ctgacgtc
//
