LOCUS       sequence-405392-        7158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GpACR1 in fusion with mScarlet in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7158)
  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 7158)
  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: GpACR1-pcDNA3.1-mScarlet
FEATURES             Location/Qualifiers
     source          1..7158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     554..573
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        554..572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1679..2374
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1812..1830)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2080..2099
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2440..2457)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2446..2670
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2716..3144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2803..2822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3013..3034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3153..3173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3158..3487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3338..3473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3400..3419
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3554..4348
                     /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(3608..3627)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4218..4237
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4522..4643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4559..4578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4613..4632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4692..4708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4692..4708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4705..4727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4716..4732
                     /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(4740..4746,4747..4764,4765..4770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4785..4806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4923..4940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5094..5682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5174..5193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5853..6644,6645..6713))
                     /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     6476..6495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6714..6818)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6893..6912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7084..7158,1..305)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      541 ggcttatcga aattaatacg actcactata gggagaccca agctggctag cgtttaaacg
      601 ggccctctag actcgagcgg ccgccactgt gctggatatc tgcagaattc caccacactg
      661 gactagtgga tccccgggta ccggtgccac catgctcgct cttgatgaag tcgccacctt
      721 tacccaatgg tctgtgatgg caataagttt tctgcagttc gcattccttg tcgacagatt
      781 taggcagggg cagtgtttcg ttgaggctct gtacgtgata atggtggaat ttatcggcta
      841 cgggttgaac gtggcagggt ggtatactgt aaacctggag gacggacgaa ccttttatat
      901 tggccgctac gtttcctgga tccttacatc ccctgtcctc cttatcatag tatgtaacct
      961 tcacgccgag gctcgcattg aacccagaag gacattgttc tgctccgtga tgtttccctt
     1021 gatgactgcc ctcgggggcg cagccgcctt gatcgaaatt atatgggtaa gagttgcact
     1081 gttccttctt agctgtgtta tctgtgggct gatttttaga gtagttcaca tatgccgaac
     1141 aaaaagtctg attgtcgtgg gcggaaacga cactgctaga cgcctcattg accgaataac
     1201 ctggcttttt tacgtgacct ggtgtatgta tccaaccttt gtgctcttgg ggcccgaatt
     1261 tttcgatgta gtgcctgccg caggagtcgc cgtctgggcc gcagtcactg atcttttcag
     1321 caaaaacctg ttcggattgt tgtgttggtt tcttcggtgg caggttattg aacccagagg
     1381 taggagagtc gtggggcttg tgaagcaatt cagtcggctg aattcaatgc tcaataatgt
     1441 cgaagatacc cccaggccaa gcccacacgg caagcacgta gttttgaata ccggcagtgc
     1501 ttttgctgac actcctgagg gagtgcagtt gaccgcccca agtccttctc catcaccacg
     1561 ccttggatct ttgccccctc cagctctgcc catgcctgcg gcaccggtga agagcaggat
     1621 caccagcgag ggcgagtaca tccccctgga ccagatcgac atcaacgtgg cggtacccat
     1681 ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg
     1741 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1801 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat
     1861 cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc
     1921 cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1981 ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta
     2041 caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac
     2101 aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga
     2161 cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac
     2221 ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga
     2281 catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg
     2341 ccactccacc ggcggcatgg acgagctgta caagttctgc tacgagaacg aggtgtaatg
     2401 agaattcgat atcaagctta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt
     2461 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2521 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2581 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2641 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2701 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2761 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2821 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2881 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2941 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3001 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3061 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3121 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3241 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3301 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3361 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3421 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3481 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3541 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3601 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3661 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3721 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3781 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3841 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3901 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3961 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4021 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4081 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4141 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4201 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4261 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4321 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4381 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4441 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4501 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4561 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4621 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4681 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4741 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4801 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4861 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4921 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4981 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5041 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5101 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5161 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5221 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5281 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5341 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5401 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5461 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5521 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5581 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5641 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5701 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5761 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5821 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5881 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5941 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6001 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6061 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6121 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6181 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6241 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6301 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6361 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6421 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6481 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6541 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6601 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6661 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6721 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6781 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6841 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6901 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6961 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7021 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     7081 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     7141 gcccatatat ggagttcc
//
