LOCUS       sequence-86377-e        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-Dependent Chloride Channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7317)
  AUTHORS   Berndt A, Lee SY, Ramakrishnan C, Deisseroth K
  TITLE     Structure-guided transformation of channelrhodopsin into a 
            light-activated chloride channel.
  JOURNAL   Science. 2014 Apr 25;344(6182):420-4. doi: 10.1126/science.1252367.
  PUBMED    24763591
REFERENCE   2  (bases 1 to 7317)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2014-04-25- 25"; volume: "344"; 
            issue: "6182"; pages: "420-4"
COMMENT     Sequence Label: pAAV-Ef1a-DIO SwiChRca-TS-EYFP
FEATURES             Location/Qualifiers
     source          1..7317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1450..1483)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1534..1567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1579..2292,2293..2295))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFREDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1624..1645)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1973..1992
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2232..2253
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3288..3297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3302..3335
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3386..3419
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3444..4032
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3497..3517)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3915..3926)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4064..4540
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4263..4282)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4580..4720
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4580..4709
                     /label=AAV2 ITR
     rep_origin      4795..5250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4882..4901)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5092..5113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5267..5286)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5386..5408
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5446..5464)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5532..5636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5637..5705,5706..6497)
                     /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(5855..5874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6668..7256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7157..7176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccctatt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1621 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     1681 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1741 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     1801 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     1861 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccctgaagt cgatgccctt
     1921 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1981 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     2041 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta
     2101 gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     2161 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     2221 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     2281 ctcgcccttg ctcaccacgt tgatgtcgat ctggtccagg gggatgtact cgccctcgct
     2341 ggtgatcctg ctcttggcgg ccgcgaccgc gccagcctcg gcctcgtcct ccaccagcgt
     2401 ctcgacctca atctcagtgc caccaatgtt caatttggtg gtcttgcgaa tgtcgccgtg
     2461 gatgaggata tgctcgtgga tcagcacgcg caggtagtgg ccgagcagac cccagcactg
     2521 cttcgacatc aggtcaatga tggtgtggcc gacgttggag ccgtacttgc tcaggacgcc
     2581 gaagccctcg gggccgagga tgaacaggat ggggaacata ccccatgata cgaagaagag
     2641 ccaagccatg ccagtcacca cctggcgaca ccggcccttc ggcacggtgt ggtacgcctc
     2701 aatgtagacc ttggctgcgt tgaagaatgt gtagatgccg tagcacaggc ccatcaggaa
     2761 gaaaatgaca cggacgtatc ccttggacag cgcggccgtg gtgccccaca cgatcgtgcc
     2821 gatatctgac accagcagac ccatggtacg cttgttatag tcgttcgcca gacccgtaag
     2881 gttgctcaga cggataagaa tgacaggggc ggtcagcagc cagctcgcgt aacgaagcca
     2941 cttggtcttg ttgccgttgg atgagtagat caccgcaggt tcgtcaaagc tatggaaata
     3001 ctcgatgatg aacttgatca tcgagatcgt ggccacgtaa atctcctccc agccgcaagt
     3061 agacttccag gtctggtagc cgtagaacat caggcagagc gctgaaagcg caaaagaaat
     3121 ccactgcaga atgttggcag ccaacttctc ggcatttgtt ccgttggatt taagccaagc
     3181 caagcagaag cactggccgt tgttcgggat gcaaataaca gaaccattgt tctcaagagt
     3241 gtatgaggtt tggaagagtc cgacggcaga caaagcgccg ccatagtcca tggtggctag
     3301 cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact
     3361 gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg
     3421 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3481 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3541 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3601 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3661 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3721 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3781 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3841 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     3901 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3961 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4021 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     4081 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4141 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4201 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4261 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4321 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4381 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     4441 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4501 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     4561 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4621 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4681 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4741 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4801 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4861 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4921 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4981 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5041 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5101 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5161 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5221 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5281 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5341 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5401 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5461 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5521 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5581 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5641 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5701 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5761 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5821 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5881 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5941 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6001 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6061 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6121 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6181 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6241 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6301 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6361 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6421 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6481 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6541 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6781 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
