LOCUS       sequence-199318-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anion channelrhodopsin from unclassified cryptophyte (CCMP2293) for 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  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 7086)
  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"; journalName: "Sci Rep."; 
            date: "2017-03-3"; volume: "7:43358. doi"; pages: " 
            10.1038/srep43358"
COMMENT     Sequence Label: pcDNA3.1-C1ACR_023-EYFP
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(242..670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(352..373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     564..583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    716..940
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     929..946
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(969..986)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(996..1037)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(join(1067..1780,1781..1783))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1112..1133)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1461..1480
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1720..1741
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2739..2758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2740..2758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2803..3006)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2832..2852)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3007..3386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3558..3577
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3652..3756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3757..3825,3826..4617)
                     /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     complement(3975..3994)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4788..5376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5277..5296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5530..5547
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5664..5685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5700..5705,5706..5723,5724..5730)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5738..5754
                     /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)."
     primer_bind     5743..5765
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5762..5778
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5762..5778
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5827..5948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5838..5857)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5892..5911
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6124..6918)
                     /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(6235..6254)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6845..6864
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6985..7086,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6999..7086,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7053..7072)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
      241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
      361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
      421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
      481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
      541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
      601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
      661 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
      721 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
      781 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
      841 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
      901 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
      961 taaactcaat ggtgatggtg atgatgaccg gtacgcgtag aatcgagacc gaggagaggg
     1021 ttagggatag gcttaccttc gaaccgcggg ccctctagag catatgttac ttgtacagct
     1081 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     1141 gcttctcgtt ggggtctttg ctcagggcgg actggtagct caggtagtgg ttgtcgggca
     1201 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     1261 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     1321 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     1381 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     1441 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     1501 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     1561 ggtagcgggc gaagcactgc aggccgtagc cgaaggtggt cacgagggtg ggccagggca
     1621 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     1681 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     1741 ggatgggcac caccccggtg aacagctcct cgcccttgct cacggcggcc gccctcacgc
     1801 tgccgatcac cttgccgaac atcttcacct ccacgtcgct cttgggtctg ggggcgtcct
     1861 gcacgtcctt ctcggattcg gcagcaggag gggccctcac gaagtcccta tcccacttgc
     1921 cgttcagcag gccccaggtg gtggaccaca gcaggattcc gttgaagttc ttgcagaagg
     1981 cgtcgctaat caggtaggcc acggcgctga tgttctcgtc gatcacgcat gtgccggtgc
     2041 tgctcaggag ccacagaatg gggaaggcgg tccaggcgac gaagaagatg ctcctgatgc
     2101 tcttcagcct ggccaccacc tggttgccca gatcggtctt ggcgtcggca aagtcgccaa
     2161 tggtcacggc gaagatggtg tacacgctgg cgtactcgaa cagcaggcag accagtccgc
     2221 agccgaagaa ggcccacttg aggtcttcgg tcaccacgat ggtagcggtc acgcccatca
     2281 ccactctgat gatgctggcg gcgatgacca tggggttgat agggatgctc tggtacttga
     2341 tcagggtcat gccggagatc tgtccgtgca tcacggggca ggtcaccagc cagctggcca
     2401 tcctggccca aggcagcact ctgccggagc ccatcttcag gtagccgttt cctgtgtagg
     2461 ccatgacgta gacgatggcc tccaccatgg gcaggtagat ggcttcccag ggcaccttgg
     2521 gcattctggt catcatgaag gcggactggc acatgctcac cacggcccag cagatctgaa
     2581 acaccttcac cacgggcacg agctcggcag ggatacactg gggatgggag aagatgtatt
     2641 cgccgtacat cactccgtcg atgatcacgg cttcgttcac ttccagcacg gtcatggatc
     2701 cgagctcggt accaagctta actagccagc ttgggtctcc ctatagtgag tcgtattaat
     2761 ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac
     2821 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2881 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2941 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     3001 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     3061 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     3121 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     3181 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     3241 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     3301 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     3361 ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag
     3421 cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt
     3481 tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag
     3541 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc
     3601 gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3661 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3721 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3781 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3841 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3901 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3961 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4021 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4081 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4141 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4201 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4261 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4321 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4381 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4441 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4501 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4561 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4621 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4681 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4741 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4801 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4861 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4921 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4981 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5041 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5101 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5161 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5221 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5281 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5341 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5401 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5461 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5521 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5581 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5641 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5701 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5761 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag
     5821 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     5881 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5941 aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     6001 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     6061 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgcgaac cccagagtcc
     6121 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     6181 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     6241 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     6301 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     6361 cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg
     6421 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     6481 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     6541 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     6601 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     6661 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     6721 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     6781 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     6841 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     6901 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atccgaaaat
     6961 ggatatacaa gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     7021 ctacttctgg aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat
     7081 tagtca
//
