LOCUS       Exported                7083 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Anion channelrhodopsin from Rhodomonas sp. (CCMP768) for expression 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-R1ACR_877-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Govorunova EG, Sineshchekov OA, Rodarte EM, Janz R, Morelle O, 
            Melkonian M, Wong GK, Spudich JL
  TITLE     The Expanding Family of Natural Anion Channelrhodopsins Reveals 
            Large Variations in Kinetics, Conductance, and Spectral Sensitivity.
  JOURNAL   Sci Rep. 2017 Mar 3;7:43358. doi: 10.1038/srep43358.
  PUBMED    28256618
REFERENCE   2  (bases 1 to 7083)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7083
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(13..33)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        18..347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      198..333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     260..279
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             414..1208
                     /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(468..487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1078..1097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1384..1505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1421..1440)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1475..1494
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1554..1570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1554..1570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1567..1589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1578..1594
                     /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(1602..1632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1647..1668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1785..1802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1956..2544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2036..2055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2715..3575)
                     /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     3338..3357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3576..3680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3755..3774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3946..4325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4326..4529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4480..4500
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4574..4593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4574..4592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5546..6262
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5588..5609)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5849..5868)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6196..6217
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6292..6333
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6343..6360
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6383..6400)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6389..6613
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6659..4
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6746..6765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6956..6977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
       61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
      181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
      301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
      361 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
      421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
      481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
      541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
      601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
      661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
      721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
      781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
      841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
      901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
      961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     1021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     1081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     1141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     1201 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     1261 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1321 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1381 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1441 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1501 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     1561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1741 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2641 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2701 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2761 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2821 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2881 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2941 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3001 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3061 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3121 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3181 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3241 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3301 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3361 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3421 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3481 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3541 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3601 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3661 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     3721 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3781 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     3841 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     3901 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     3961 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4021 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4081 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4141 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4201 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4261 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4321 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4381 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4441 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4501 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     4561 ggcttatcga aattaatacg actcactata gggagaccca agctggctag ttaagcttgg
     4621 taccgagctc ggatccatga gcgccaccga aaaccctgcc ttcggcctgt acggcacctg
     4681 ggagaacgac cacccccagt gtgtggagat gggcctggac accagcagcc agacaaacat
     4741 gatccacctc ctgttcgcca gcatctgctt ctgtcaggtg ctctttatgg cctccagagc
     4801 tcccagagtg acctgggagg ccgtctacct gcccctggtg gagaccgtgc tgtacggaat
     4861 ggccgccagc ggcaacggca acctgaggat ctccaacggc aggatggtgc ccatggccag
     4921 gttctgcagc tggctgatca cctgtcctat catgctgctg caggtggtgg tgatgcacga
     4981 cgtggtgtgg ttcaacgtga gcaccaggta catggtgctg gccgccagcc tgatcaggac
     5041 cgtgttcggc atcggcgcca gcgtgaccac caacgatacc atgagatggg tgcagttcct
     5101 gctgtcctgc atctgcttcg ccttcgagat ggtggtggcc caccagacct acaagggagc
     5161 cctgaagaag tttgccgccg tgaagaccac cctgaacggc gtggtctacg ccaggatcat
     5221 gtttctgagg ttcctgttct tcttcagctg gaacgccttc cctatcatct gggccctgag
     5281 cagcacaggc ctgtgtctga tcagcgagga cgccagcgtg ctgtcctatc tcgtggccga
     5341 cctgatctgc aagaacatct acggcatcac caacgccaac accgtgttca gagtcctgga
     5401 cggagcctgg gtccctgaga ccgaggaaca cgcccaggcc aaggaccacc tgacctacct
     5461 cgagatgatg atgaaccaca acctgggcat cggcagacat catgacgagg agctgcaaat
     5521 ggaagtgggc catgacgcgg ccgccgtgag caagggcgag gagctgttca ccggggtggt
     5581 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5641 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5701 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc
     5761 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5821 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5881 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5941 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     6001 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     6061 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     6121 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     6181 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     6241 catggacgag ctgtacaagt aacatatgct ctagagggcc cgcggttcga aggtaagcct
     6301 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat
     6361 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6421 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6481 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6541 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6601 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     6661 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6721 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6781 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6841 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6901 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6961 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7021 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7081 gcg
//
