LOCUS       Exported                7054 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses channelrhodopsin-EGFP fusion in CamKIIa+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-CamKIIa-ChR2-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  AUTHORS   Keaveney MK, Tseng HA, Ta TL, Gritton HJ, Man HY, Han X
  TITLE     A MicroRNA-Based Gene-Targeting Tool for Virally Labeling 
            Interneurons in the Rodent Cortex.
  JOURNAL   Cell Rep. 2018 Jul 10;24(2):294-303. doi: 
            10.1016/j.celrep.2018.06.049.
  PUBMED    29996091
REFERENCE   2  (bases 1 to 7054)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7054
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   2..131
                     /label=AAV2 ITR
     promoter        198..1486
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1500..1509
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1506..2432
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             2454..3170
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYTYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2496..2517)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2757..2776)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3104..3125
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3210..3798
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3263..3283)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3681..3692)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3801..3817)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3802..3818)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3847..3968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3858..3877)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3912..3931
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4278..4295)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4278..4294)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4434..4889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4521..4540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4731..4752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4915..5019
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5020..5880
                     /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(5238..5257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6051..6639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6540..6559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6793..6810
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6927..6948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6963..6993
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7001..7017
                     /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     7006..7028
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7025..7041
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7025..7041
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      181 taggaagatc tgaattccat tatggcctta ggtcacttca tctccatggg gttcttcttc
      241 tgattttcta gaaaatgaga tgggggtgca gagagcttcc tcagtgacct gcccagggtc
      301 acatcagaaa tgtcagagct agaacttgaa ctcagattac taatcttaaa ttccatgcct
      361 tgggggcatg caagtacgat atacagaagg agtgaactca ttagggcaga tgaccaatga
      421 gtttaggaaa gaagagtcca gggcagggta catctacacc acccgcccag ccctgggtga
      481 gtccagccac gttcacctca ttatagttgc ctctctccag tcctaccttg acgggaagca
      541 caagcagaaa ctgggacagg agccccagga gaccaaatct tcatggtccc tctgggagga
      601 tgggtgggga gagctgtggc agaggcctca ggaggggccc tgctgctcag tggtgacaga
      661 taggggtgag aaagcagaca gagtcattcc gtcagcattc tgggtctgtt tggtacttct
      721 tctcacgcta aggtggcggt gtgatatgca caatggctaa aaagcaggga gagctggaaa
      781 gaaacaagga cagagacaga ggccaagtca accagaccaa ttcccagagg aagcaaagaa
      841 accattacag agactacaag ggggaaggga aggagagatg aattagcttc ccctgtaaac
      901 cttagaaccc agctgttgcc agggcaacgg ggcaatacct gtctcttcag aggagatgaa
      961 gttgccaggg taactacatc ctgtctttct caaggaccat cccagaatgt ggcacccact
     1021 agccgttacc atagcaactg cctctttgcc ccacttaatc ccatcccgtc tgttaaaagg
     1081 gccctatagt tggaggtggg ggaggtagga agagcgatga tcacttgtgg actaagtttg
     1141 ttcgcatccc cttctccaac cccctcagta catcaccctg ggggaacagg gtccacttgc
     1201 tcctgggccc acacagtcct gcagtattgt gtatataagg ccagggcaaa gaggagcagg
     1261 ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta ccggggcaac gggaacaggg
     1321 cgtttcggag gtggttgcca tggggacctg gatgctgacg aaggctcgcg aggctgtgag
     1381 cagccacagt gccctgctca gaagccccaa gctcgtcagt caagccggtt ctccgtttgc
     1441 actcaggagc acgggcaggc gagtggcccc tagttctggg ggcagcgggg cgcgccgccg
     1501 ccaccatgga ctatggcggc gctttgtctg ccgtcggacg cgaacttttg ttcgttacta
     1561 atcctgtggt ggtgaacggg tccgtcctgg tccctgagga tcaatgttac tgtgccggat
     1621 ggattgaatc tcgcggcacg aacggcgctc agaccgcgtc aaatgtcctg cagtggcttg
     1681 cagcaggatt cagcattttg ctgctgatgt tctatgccta ccaaacctgg aaatctacat
     1741 gcggctggga ggagatctat gtgtgcgcca ttgaaatggt taaggtgatt ctcgagttct
     1801 tttttgagtt taagaatccc tctatgctct accttgccac aggacaccgg gtgcagtggc
     1861 tgcgctatgc agagtggctg ctcacttgtc ctgtcatcct tatccacctg agcaacctca
     1921 ccggcctgag caacgactac agcaggagaa ccatgggact ccttgtctca gacatcggga
     1981 ctatcgtgtg gggggctacc agcgccatgg caaccggcta tgttaaagtc atcttctttt
     2041 gtcttggatt gtgctatggc gcgaacacat tttttcacgc cgccaaagca tatatcgagg
     2101 gttatcatac tgtgccaaag ggtcggtgcc gccaggtcgt gaccggcatg gcatggctgt
     2161 ttttcgtgag ctggggtatg ttcccaattc tcttcatttt ggggcccgaa ggttttggcg
     2221 tcctgagcgt ctatggctcc accgtaggtc acacgattat tgatctgatg agtaaaaatt
     2281 gttgggggtt gttgggacac tacctgcgcg tcctgatcca cgagcacata ttgattcacg
     2341 gagatatccg caaaaccacc aaactgaaca tcggcggaac ggagatcgag gtcgagactc
     2401 tcgtcgaaga cgaagccgag gccggagccg tgccagcggc accggtagta gcagtgagca
     2461 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2521 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2581 ccctgaagtt catttgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2641 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2701 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2761 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2821 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2881 acacctacaa cagccacaat gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2941 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3001 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3061 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3121 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa actagtgcgg
     3181 ccgccctgca ggcatgcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3241 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3301 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3361 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3421 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3481 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3541 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3601 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3661 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3721 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3781 tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg cttcgagcag
     3841 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3901 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3961 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     4021 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     4081 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     4141 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgcccgggct
     4201 ttgcccgggc ggcctcagtg cgttgcccst gcggcctcag tgckgcagak rgcggccmgt
     4261 gngaggtatt agcactcact ggccgtcgtt ttacaacgtc gtgaagggcg cccsccgccc
     4321 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctggcgta atagcgaaga
     4381 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     4441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4501 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4561 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4621 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4741 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4801 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4861 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     4921 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4981 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5041 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5101 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5161 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5221 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5281 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5341 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5401 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5461 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5521 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5581 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5641 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5701 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5761 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5821 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5881 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5941 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6001 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6121 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6181 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6241 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6301 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6361 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6421 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6481 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6541 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6601 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6661 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6721 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6781 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6901 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6961 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7021 cacacaggaa acagctatga ccatgattac gcca
//
