LOCUS       Exported                7079 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 7079)
  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 7079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7079
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        197..1485
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1499..1508
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1505..2431
                     /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             2453..3166
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2495..2516)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2756..2775)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3103..3124
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3209..3797
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3262..3282)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3680..3691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3800..3816)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3801..3817)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3846..3967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3857..3876)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3911..3930
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4129..4258
                     /label=AAV2 ITR
     primer_bind     complement(4277..4294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4277..4293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4286..4308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4435..4890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4522..4541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4732..4753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4916..5020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5021..5881
                     /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(5239..5258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6052..6640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6541..6560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6794..6811
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6928..6949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6964..6994
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7002..7018
                     /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     7007..7029
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7026..7042
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7026..7042
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct gaattccatt atggccttag gtcacttcat ctccatgggg ttcttcttct
      241 gattttctag aaaatgagat gggggtgcag agagcttcct cagtgacctg cccagggtca
      301 catcagaaat gtcagagcta gaacttgaac tcagattact aatcttaaat tccatgcctt
      361 gggggcatgc aagtacgata tacagaagga gtgaactcat tagggcagat gaccaatgag
      421 tttaggaaag aagagtccag ggcagggtac atctacacca cccgcccagc cctgggtgag
      481 tccagccacg ttcacctcat tatagttgcc tctctccagt cctaccttga cgggaagcac
      541 aagcagaaac tgggacagga gccccaggag accaaatctt catggtccct ctgggaggat
      601 gggtggggag agctgtggca gaggcctcag gaggggccct gctgctcagt ggtgacagat
      661 aggggtgaga aagcagacag agtcattccg tcagcattct gggtctgttt ggtacttctt
      721 ctcacgctaa ggtggcggtg tgatatgcac aatggctaaa aagcagggag agctggaaag
      781 aaacaaggac agagacagag gccaagtcaa ccagaccaat tcccagagga agcaaagaaa
      841 ccattacaga gactacaagg gggaagggaa ggagagatga attagcttcc cctgtaaacc
      901 ttagaaccca gctgttgcca gggcaacggg gcaatacctg tctcttcaga ggagatgaag
      961 ttgccagggt aactacatcc tgtctttctc aaggaccatc ccagaatgtg gcacccacta
     1021 gccgttacca tagcaactgc ctctttgccc cacttaatcc catcccgtct gttaaaaggg
     1081 ccctatagtt ggaggtgggg gaggtaggaa gagcgatgat cacttgtgga ctaagtttgt
     1141 tcgcatcccc ttctccaacc ccctcagtac atcaccctgg gggaacaggg tccacttgct
     1201 cctgggccca cacagtcctg cagtattgtg tatataaggc cagggcaaag aggagcaggt
     1261 tttaaagtga aaggcaggca ggtgttgggg aggcagttac cggggcaacg ggaacagggc
     1321 gtttcggagg tggttgccat ggggacctgg atgctgacga aggctcgcga ggctgtgagc
     1381 agccacagtg ccctgctcag aagccccaag ctcgtcagtc aagccggttc tccgtttgca
     1441 ctcaggagca cgggcaggcg agtggcccct agttctgggg gcagcggggc gcgccgccgc
     1501 caccatggac tatggcggcg ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa
     1561 tcctgtggtg gtgaacgggt ccgtcctggt ccctgaggat caatgttact gtgccggatg
     1621 gattgaatct cgcggcacga acggcgctca gaccgcgtca aatgtcctgc agtggcttgc
     1681 agcaggattc agcattttgc tgctgatgtt ctatgcctac caaacctgga aatctacatg
     1741 cggctgggag gagatctatg tgtgcgccat tgaaatggtt aaggtgattc tcgagttctt
     1801 ttttgagttt aagaatccct ctatgctcta ccttgccaca ggacaccggg tgcagtggct
     1861 gcgctatgca gagtggctgc tcacttgtcc tgtcatcctt atccacctga gcaacctcac
     1921 cggcctgagc aacgactaca gcaggagaac catgggactc cttgtctcag acatcgggac
     1981 tatcgtgtgg ggggctacca gcgccatggc aaccggctat gttaaagtca tcttcttttg
     2041 tcttggattg tgctatggcg cgaacacatt ttttcacgcc gccaaagcat atatcgaggg
     2101 ttatcatact gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg catggctgtt
     2161 tttcgtgagc tggggtatgt tcccaattct cttcattttg gggcccgaag gttttggcgt
     2221 cctgagcgtc tatggctcca ccgtaggtca cacgattatt gatctgatga gtaaaaattg
     2281 ttgggggttg ttgggacact acctgcgcgt cctgatccac gagcacatat tgattcacgg
     2341 agatatccgc aaaaccacca aactgaacat cggcggaacg gagatcgagg tcgagactct
     2401 cgtcgaagac gaagccgagg ccggagccgt gccagcggca ccggtagtag cagtgagcaa
     2461 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2521 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2581 cctgaagttc atttgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2641 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2701 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2761 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2821 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2881 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2941 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3001 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3061 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3121 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa ctagtgcggc
     3181 cgccctgcag gcatgcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3241 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3301 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3361 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3421 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3481 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3541 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3601 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3661 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3721 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3781 ttgggccgcc tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga
     3841 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3901 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3961 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4021 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     4081 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     4141 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4201 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc
     4261 ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4321 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4381 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     4441 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4501 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4561 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4621 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4681 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4741 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4801 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4861 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4981 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5401 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5581 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6661 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6721 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6781 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6901 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6961 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7021 tcacacagga aacagctatg accatgatta cgccagattt aattaaggcc ttaattagg
//
