LOCUS       Exported                7170 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Anion channelrhodopsin from unclassified cryptophyte (CCMP2293) for 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-C1ACR_887-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7170)
  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 7170)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7170
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(135..154)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     482..503
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             578..619
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             629..646
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(669..686)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    675..899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      945..1373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1032..1051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1242..1263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1382..1402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1387..1716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1567..1702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1629..1648
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1783..2577
                     /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(1837..1856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2447..2466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2753..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2844..2863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2923..2939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2923..2939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2936..2958)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2947..2963
                     /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(2971..3001)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3016..3037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3154..3171)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3325..3913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3405..3424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4084..4944)
                     /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     4707..4726
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4945..5049)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5124..5143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5315..5694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5695..5898
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5849..5869
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5943..5962
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5943..5961
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7002..548
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(7044..7065)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc
       61 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      121 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      181 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      241 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      301 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      361 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      421 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      481 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      541 acaagtaaca tatgctctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
      601 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
      661 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
      721 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
      781 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
      841 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
      901 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
      961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     1381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     1501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1621 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     1681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     1741 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     1801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     1861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     1921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     1981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     2221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     2281 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     2341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     2401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     2461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     2521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     2581 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     2641 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2701 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     2761 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2821 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2881 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     2941 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3001 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3061 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3121 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3181 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3241 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3301 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3361 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3421 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3481 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3541 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3661 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3721 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3781 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3841 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3961 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4021 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4081 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4201 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4381 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4681 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5041 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     5101 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     5161 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     5221 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     5281 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     5341 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5401 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5461 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5521 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5581 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5641 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     5701 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     5761 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     5821 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     5881 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     5941 attaatacga ctcactatag ggagacccaa gctggctagt taagcttggt accgagctcg
     6001 gatccatgga gcatcacagc tacgatggcg ccatgaggag cctgcaggct atcagcggct
     6061 ccttcgccga catcgaggga cactggaacg gccccacctc cgtgctggat aaccccgccg
     6121 tgatcgacgg acagctgtac ggagcctatg tcttcatgca ccctgagtgc caccagggcg
     6181 agtacgcccc cattgtgaag atcatccagc tgtgctgggc cgtgatctgc atgtgccaag
     6241 tgctgttcat gttcagcagg gcccctaggg tgccctggga ggccatgtat ctgcccggcg
     6301 tggagtgcgt gatctacacc ctggccttca caggcaacgg ctacgtgagg atggtggacg
     6361 gcagaatcct gacctggagc agactggcca gctggctggt gtgttgccct gtgatgctgg
     6421 gccagatctc cgccatcacc ctgatcaagt accagtccat ccctctgaac cccatgatga
     6481 tcgcctgcag catcattagg accgtgtttg gcgtgagcgc tggcgtggtg caagaggagc
     6541 ccctgaagtg ggtgttcttc tggctgggca tcttcttttt cgccttcgag ctgctgggct
     6601 gctacctgat tttcgctatc gccatcgccg acttcgaggc ctgcaagacc gaactggcca
     6661 acaaagtgat cggcagaatc aagaccatga gggccatcat cttcatctgc tggcccgcca
     6721 tcaccgtgac atgggtgctg tcctccgccg gcacctgtct ggtggacgac aactacacca
     6781 ccatcggctt cgtcgtgctg gacgctgtgt gcaagaacat gtacggcgtc gtcatctgga
     6841 gcaccacctg gggcctcctg aacggcaagt gggacagaga ctacgccaga accttcggcc
     6901 aggaggaaga acagcaggag gaagcccctg actttgacgc caagggagga gccatgcctg
     6961 gcgccaacga caatgtggag gtgaagatgt tcgcggccgc cgtgagcaag ggcgaggagc
     7021 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     7081 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     7141 tctgcaccac cggcaagctg cccgtgccct
//
