LOCUS       sequence-300511-        5761 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Anion-conducting channelrhodopsin of viral origin. Codon-optimized 
            for mammalian expression (human/mouse)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5761)
  AUTHORS   Rozenberg A, Oppermann J, Wietek J, Fernandez Lahore RG, Sandaa RA, 
            Bratbak G, Hegemann P, Beja O
  TITLE     Lateral Gene Transfer of Anion-Conducting Channelrhodopsins between 
            Green Algae and Giant Viruses.
  JOURNAL   Curr Biol. 2020 Dec 21;30(24):4910-4920.e5. doi: 
            10.1016/j.cub.2020.09.056. Epub 2020 Oct 15.
  PUBMED    33065010
REFERENCE   2  (bases 1 to 5761)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cub.2020.09.056"; journalName: "Curr Biol"; date: 
            "2020-12-21- 21"; volume: "30"; issue: "24"; pages: "4910-4920.e5"
COMMENT     Sequence Label: vPyACR_2164382
FEATURES             Location/Qualifiers
     source          1..5761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    652..708
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      709..718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1744..1746,1747..2457)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1786..1807)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2047..2066)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2394..2415
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2629..2648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2683..2702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2720..3175)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2857..2878)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3069..3088
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3202..3306
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3308..3665
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3308..3499
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3331..3351)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3516..3651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3700..4494
                     /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             3700..4494
                     /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(3754..3773)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4364..4383
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4682..4701)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4726..4773
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5102..5690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5591..5610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
      661 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccgc caccatggac
      721 tatgggggag cactgtccgc cgtcgggctg tttcagactt cctacaccct ggagaacaac
      781 gggagcgtca tttgcattcc taacaatggc cagtgcttct gtctggcctg gctgaagagc
      841 aacgggatgg tcagcgacac ccgaatcagg gagacactgc aggtcatcga tctgctgtgg
      901 tccatctcac tgtttgcttt ctttatgtat cagaagatgt gcaagaacag gtgtaagttt
      961 gaagtgatgt acgtgagctc catcggctcc atcaccgacc tggtgaagct ggccttcacc
     1021 gattatacac ctgcctctat cacaatcaag gagaacgaca tcgtgtttaa ttggagccag
     1081 tactatggct ggctgatcac ctgcccagtg ctgctgatcc acctgtctaa tctggccggc
     1141 aaggacgtgt ttgatgtgag gagaatgatg aagatcctgg tggcctacca gatcctgatg
     1201 ctgtccggcg ccacagcctc cctgtctgag aaggattctc ccgtgaagtg gtgcttcttt
     1261 atcctggcca tcagctgtct gatgatcctg taccgctacg ccgcccacat cttcctggag
     1321 agcatgaaga tcatgcccaa gaacgccacc tccgtgctga agcagatcgc catcgtgttc
     1381 tactctagct ggagcggctt tggcttcttt tggatcatcg gaccttccgg cctggccctg
     1441 gtgagcaacg agatctccaa ggccggcttc gccttttgcg acatcctgtc caagaatatc
     1501 tattctatgc tgggctggta tctgaggtgg tatgtgctga gaaagcacga caagccagat
     1561 gagttcatca accagaccga gacacccctg aagaagatca ccatcctgct ggtggataac
     1621 gagcacaatt tcctgcacta cttttccaca atcctgtatc agctggactg cgatatctac
     1681 gtgaagagca ggatcaccag cgagggcgag tacatccccc tggaccagat cgacatcaac
     1741 gtggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1801 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1861 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1921 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     1981 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2041 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2101 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2161 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2221 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2281 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2341 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2401 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagttc
     2461 tgctacgaga acgaggtgtg atctagatca taatcagcca taccacattt gtagaggttt
     2521 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2581 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2641 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2701 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2761 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2821 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2881 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2941 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3001 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3061 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3121 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3181 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3241 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3301 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3481 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3601 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3661 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3721 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3781 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3841 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     3901 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3961 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4021 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4081 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4141 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4201 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4261 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4321 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4381 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4441 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4501 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4561 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4621 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     4681 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4741 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4801 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4861 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4921 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     4981 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5041 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5221 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5761 c
//
