LOCUS       sequence-405398-        5427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the MsACR1 mutant raACR as a fusion protein with 
            mCerulean3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5427)
  AUTHORS   Oppermann J, Rozenberg A, Fabrin T, Gonzalez-Cabrera C, Parker R, 
            Beja O, Prigge M, Hegemann P
  TITLE     Robust optogenetic inhibition with red-light-sensitive 
            anion-conducting channelrhodopsins.
  JOURNAL   eLife. 2024 Oct 14;12:RP90100. doi: 10.7554/eLife.90100.
  PUBMED    39401075
REFERENCE   2  (bases 1 to 5427)
  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: "2024-10-14"; volume: "12:RP90100. doi"; pages: " 
            10.7554/eLife.90100"
COMMENT     Sequence Label: raACR-pmCerulean3-N1
FEATURES             Location/Qualifiers
     source          1..5427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     466..486
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1271..1273,1274..1987)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIKA
                     NFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1313..1334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1574..1593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1921..1942
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2162..2283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2199..2218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2253..2272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2290..2745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2427..2448)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2639..2658
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2772..2876
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2878..3235
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2878..3069
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2901..2921)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3086..3221
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3270..4064
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3270..4064
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3324..3343)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3934..3953
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4252..4271)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4296..4343
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4672..5260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5161..5180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      541 agccaccatg tccgcctacg aggtggtgaa ggacatccgg ctggtcatcc agaccatcct
      601 gctgctggcc ttttatgtga tggccggctt ctacctgtat cagaagaaga agaagagggc
      661 cagatgggag gtgctgtaca tcgcaaccgt ggcaggcctg gccaacacaa tcggcctggc
      721 cgcaggacac ctggagccac tgtcttttgc cgcagaggga gtgggcaaga agacaccatg
      781 gctgcggttc agctcctgga tgaccacatg ccccgtgctg ctgatccacc tgaacaatct
      841 ggcaggagag gaggtgcaga acgtggagcg cgccagcatg ctggtcatcg agctgaatct
      901 gatggtgctg ctgggcgcca cctctagctt cacagatggc ggcggcgccc tggtgttctt
      961 tatgctgagc tggctggtgt gctttaccat cttcgtgcag gcctggacaa tctttaacga
     1021 ggccttcgcc tccctgccag caaaggcaag gcctcacctg aatggcatga tgttctgttt
     1081 ttacttcgcc tgggcaggat ttggcgtgct gttcgcagca ggaccagagg gatggggacc
     1141 caaggacaat atcctggccg agctggtgac catcgccatg gtggtggtgg atttcatcgc
     1201 caagatcgtg tttggcttca tcggatggca cctgaggtgg cgcgtgctga ggggagagga
     1261 cggcaaggtg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1321 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1381 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1441 gcccaccctc gtgaccaccc tgagctgggg cgtgcagtgc ttcgcccgct accccgacca
     1501 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1561 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1621 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1681 ggggcacaag ctggagtaca acgccatcca cggcaacgtc tatatcaccg ccgacaagca
     1741 gaagaacggc atcaaggcca acttcggcct caactgcaac atcgaggacg gcagcgtgca
     1801 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1861 caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     1921 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1981 caagtaggga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     2041 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2101 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2161 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2221 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2281 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2341 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2401 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2461 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2521 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2581 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2641 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2701 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2761 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2821 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2881 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2941 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3001 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3061 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3121 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3181 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3241 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3301 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3361 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3421 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3481 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3541 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3601 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3661 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3721 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3781 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3841 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3901 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3961 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4021 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4081 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4141 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4201 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4261 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4321 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4381 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4441 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4501 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4561 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4621 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4681 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4741 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4801 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4861 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4921 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4981 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5041 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5101 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5161 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5221 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5281 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5341 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     5401 ggtcattagt tcatagccca tatatgg
//
