LOCUS       sequence-199235-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anion channelrhodopsin from Geminigera sp. (Caron Lab Isolate) for 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  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 7236)
  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-G1ACR_203-EYFP
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(116..445)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      130..265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(184..203)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     430..450
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(459..887)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(569..590)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     781..800
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    933..1157
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1146..1163
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1186..1203)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1213..1254)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(join(1284..1997,1998..2000))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1329..1350)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1678..1697
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1937..1958
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3106..3125)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3107..3125)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3170..3373)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3199..3219)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3374..3753
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3925..3944
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4019..4123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4124..4192,4193..4984)
                     /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(4342..4361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5155..5743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5644..5663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5897..5914
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6031..6052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6067..6072,6073..6090,6091..6097)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6105..6121
                     /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     6110..6132
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6129..6145
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6129..6145
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6194..6315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6205..6224)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6259..6278
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6491..7236,1..49))
                     /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(6602..6621)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7212..7231
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
       61 ctcatcctgt ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc
      121 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg
      181 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
      241 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
      301 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
      361 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
      421 actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta
      481 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
      541 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
      601 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      661 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      721 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
      781 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      841 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct
      901 agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg
      961 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     1021 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     1081 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     1141 gcaactagaa ggcacagtcg aggctgatca gcgggtttaa actcaatggt gatggtgatg
     1201 atgaccggta cgcgtagaat cgagaccgag gagagggtta gggataggct taccttcgaa
     1261 ccgcgggccc tctagagcat atgttacttg tacagctcgt ccatgccgag agtgatcccg
     1321 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     1381 agggcggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     1441 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     1501 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     1561 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     1621 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     1681 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     1741 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcagg
     1801 ccgtagccga aggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     1861 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     1921 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     1981 agctcctcgc ccttgctcac ggcggccgca tcctcgatct ctttggcgga gttgaacctg
     2041 ccgtgagggg cggcgaaggt gtcttcctcc ttgtggttgt ggctgttggc gggagcctcc
     2101 tcatcgtgat tgttggggtg cttcttgggg gtccaggttc ctcccagcag gccccacacg
     2161 gtgttggtct ggatcactcc gtacaggttc ttgcacagag cgtcggcgca caggtagccg
     2221 agagcgcttg tctgttcgct gatgacgcac agtccggtgc tgctcagcac ccaaaacacg
     2281 gggaaggcgg tccagctggt aaaaaaaatg aacctcatga agtacagcct tctcagggcg
     2341 gtggcgttct caggcacatc ggtggtgggg gtaaacttgg cgatgccctg ctggaagatg
     2401 aaccacacgg tggtgatctc catggtgtaa aagctaaagc ccaggaagaa gaacagccac
     2461 ttggcgatct tggtatcggt ggttgtgctg gccagcaggc cgcacactgt cctgatgagg
     2521 gaggcggcgc aaatgggcat cctcagctgg attccgtaca gcttcatgtc gtggatgcca
     2581 aaggcctggg agagcatgat ggggcaggtg atcagccagg agcacatcct gctcaggggc
     2641 agcattctgc catcgcccat cctcagcacg ccgtatccgg ctcctgtgca gcagtacagg
     2701 atacactcca ccagaggcag gtacacggcc tcccaggtca ccttaggggc ccttgtcatc
     2761 atgaagagag cctgacagag gcacaccacg gcccacaggg taatgatggt tctctccctg
     2821 ctggtggcgt gcaggccctt gttcacgcaa tcaggattgg cgatctccca ctctccatac
     2881 atgccggcat aaggtgtggc agggccgatg ccgtcgatgt aggcggcttt gcctctcacg
     2941 accattctct cctcgggctg aaagcccacg gcctggcttc cggtatcctg agcgtccagc
     3001 atcacggcga acagcaccac cagcctggcc aggtgcacca ccttggccag cctagcggcc
     3061 atggatccga gctcggtacc aagcttaact agccagcttg ggtctcccta tagtgagtcg
     3121 tattaatttc gataagccag taagcagtgg gttctctagt tagccagaga gctctgctta
     3181 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     3241 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     3301 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     3361 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     3421 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     3481 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     3541 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     3601 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     3661 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     3721 attactatta ataactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc
     3781 gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt
     3841 gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag
     3901 ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3961 ggggatcggg agatctcccg atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg
     4021 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4081 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4141 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4201 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4261 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4321 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4381 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4441 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4501 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4561 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4621 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4681 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4741 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4801 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4861 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4921 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4981 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5041 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5101 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5821 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5881 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5941 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6001 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6061 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6121 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacgg
     6181 tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     6241 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     6301 tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     6361 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     6421 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgcgaacccc
     6481 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     6541 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     6601 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     6661 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     6721 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     6781 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     6841 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     6901 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     6961 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     7021 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     7081 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     7141 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     7201 ttgtctgttg tgcccagtca tagccgaata gcctct
//
