LOCUS       sequence-316230-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PgACR1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Govorunova EG, Sineshchekov OA, Li H, Wang Y, Brown LS, Spudich JL
  TITLE     RubyACRs, nonalgal anion channelrhodopsins with highly red-shifted 
            absorption.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Sep 15;117(37):22833-22840. doi: 
            10.1073/pnas.2005981117. Epub 2020 Sep 1.
  PUBMED    32873643
REFERENCE   2  (bases 1 to 7083)
  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"; doi: 
            "10.1073/pnas.2005981117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-09-15- 15"; volume: "117"; issue: "37"; pages: 
            "22833-22840"
COMMENT     Sequence Label: PgACR1_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        448..651
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     602..622
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     696..715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1668..1670,1671..2384)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1710..1731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1971..1990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2318..2339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2414..2455
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2465..2482
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2505..2522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2511..2735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2781..3209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2868..2887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3078..3099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3218..3238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3223..3552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3403..3538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3465..3484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3619..4413
                     /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(3673..3692)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4283..4302
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4589..4710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4626..4645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4680..4699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4759..4775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4759..4775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4772..4794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4783..4799
                     /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(4807..4813,4814..4831,4832..4837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4852..4873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4990..5007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5161..5749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5241..5260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5920..6711,6712..6780))
                     /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     6543..6562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6781..6885)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6960..6979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
       61 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      661 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      721 caagctggct agttaagctt ggtaccgagc tcggatccat ggccaacacc gacatcggcg
      781 tgaatatctt cctgctgatc gtgagcatca tggagctgct gctgctgcag atcggccaca
      841 tgagagataa ggtgaagtgg gaggccatgt gggtcatctt catcgagtgc gtggcctacg
      901 gcagctccat catctccccc gacgagggct ttgtgacaat gcacctggag aacggcatgc
      961 agatcgagtg gctgagatat gtgggctgga ttctgacctg tcctgtgctg ctgatgacac
     1021 tggtgtctat gaccacagca gagggaacca agccacctac agtgcgcctg gtgcctctgc
     1081 tggtgtgcaa tctgaccatg gtgctgttcg gcatcacatc cggctctgtg ctggagccaa
     1141 ccaagtggta cgtgtttgga atcggcgtgt gcttcggagg agtggtgttt tctagcgtgg
     1201 tgcagtgcct gtgggccctg tattgtgact ctcagaacga tagcatcaag tgcgtgagcg
     1261 ccctgctggc cctgaccttc atcacaggct ggggcatctt cccatgtaat tttgtggtgg
     1321 gccactccgg ctttggcatc gtgtctgacc agttctacat caccatgttt gtgatcggcg
     1381 atctgctgtc caagaacatc tgggtggcag tggccgtgtg gaggaaccac ctggtggacc
     1441 tggccaatgc acagcaggca gatgaggagg ccgccctgga gatgcagcag gcacagcagg
     1501 gagcagacgg caaggccgcc ctggaccaga tcgatgtgca gcggtttgag caggagtctg
     1561 tgccagattg gcggggcaat cccagaatgg cccggagagg cagctgctcc accctgatcc
     1621 tgaacgaagt gaatgccgtg acacccgaga ggaagcgcgc ggccgccgtg agcaagggcg
     1681 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1741 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1801 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     1861 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1921 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1981 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2041 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2101 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2161 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2221 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     2281 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2341 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaacatatg ctctagaggg
     2401 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     2461 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3121 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3181 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3481 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3541 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3601 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3661 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3721 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3781 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3841 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3901 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3961 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4021 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4081 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4141 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4201 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4261 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4321 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4381 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     4441 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4501 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4561 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4621 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4681 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4741 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4801 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4861 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4921 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4981 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcctt
     5161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5581 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5821 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5881 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5941 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6001 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6061 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6121 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6181 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6241 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6301 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6361 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6421 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6481 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6541 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6601 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6661 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6721 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6781 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6841 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6901 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     6961 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7021 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7081 tgc
//
