LOCUS       Exported                7101 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Anion channelrhodopsin from Rhodomonas abbreviata for expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-RaACR_687-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7101)
  TITLE     Extending the Time Domain of Neuronal Silencing with Cryptophyte 
            Anion Channelrhodopsins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7101
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1619..2335
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1661..1682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1922..1941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2269..2290
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2365..2406
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2416..2433
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2456..2473)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2462..2686
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2732..3160
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2819..2838)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3029..3050
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3169..3189)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3174..3503
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3354..3489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3416..3435
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3570..4364
                     /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(3624..3643)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4234..4253
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4540..4661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4577..4596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4631..4650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4710..4726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4710..4726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4723..4745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4734..4750
                     /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(4758..4788)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4803..4824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4941..4958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5112..5700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5192..5211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5871..6731)
                     /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     6494..6513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6732..6836)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6911..6930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc catgggcgac atgtcccaca tcgagacagt
      721 gtactatgtg aacggaaccg aggtggaccc ctggacacac aggtgccagc tgggagtgcc
      781 tggaggctcc cagggcatcc acaccttcca gctgacatgg agcctgatct gtaccgccca
      841 ggccatcttt ctgatgacaa agtggccaaa agtgggctgg gaggccatct acatgccctg
      901 ctgtgaggcc ctgtcttatg gcctgctgag cgccggacac ggatacctga agctgcacgg
      961 cgagtggcac tacatccctt ggatcagata tgccgcctgg atggtgacca caccagtgct
     1021 gctgaatcag gtgcagggcg tgctggacgt gaaagtgggc cacctgagac tggccccaat
     1081 ccagagcctg gccaacctga tcatgatcgt gatgggcgtg tccgcccacc tgcaggagaa
     1141 tatcgcctgg aagtgggtgt tctttctgat cggctgcgtg atggccctgg tcatctactg
     1201 ggtggcctgg ctggtgttct ctacccagat caagtactat gccgagacaa agacagccgt
     1261 gggcgatagg gtggtgtttc agctgaagct gatcgccgtg gccttctacg tgggctggtc
     1321 tgcctttccc ctgatctgga tgctgagctc cgagggcctg tgcgcaatcg aggagcacat
     1381 cgctgccgcc gcccacatcc ctgcagacgt gctgagcaag aacatcttcg gcctgctgat
     1441 gtggcacaca aggtggatcg tgctgggcgg ctattgggat gtgaataagt ttaaggagac
     1501 aatcgcccag cagaagctgg aggaggccaa gggcgaggag acaccccagg agaagaggaa
     1561 ggccctgatg ggacaggcag aggcctgctt caagatggcc gacgcctgtg cggccgccgt
     1621 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1681 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1741 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1801 gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1861 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1921 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1981 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2041 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2101 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2161 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2221 gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2281 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaacatat
     2341 gctctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat
     2401 tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg
     2461 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2521 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2581 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2641 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2701 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2761 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2821 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2881 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2941 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3001 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3061 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3121 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3181 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3241 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3301 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3361 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3421 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3481 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3541 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3601 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3661 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3721 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3781 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3841 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3901 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3961 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4021 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4081 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4141 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4201 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4261 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4321 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4381 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4441 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4501 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4681 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6841 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6901 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6961 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     7021 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     7081 tacgggccag atatacgcgt t
//
