LOCUS       Exported                6354 bp DNA     circular SYN 21-JUN-2022
DEFINITION  synthetic DNA.
ACCESSION   .
VERSION     .
KEYWORDS    3_pAAV-Syn_CapChR1_eYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6354)
  TITLE     NEBuilder-generated Construct
  JOURNAL   Exported  2-NOV-2020 from NEBuilder https://nebuilder.neb.com
REFERENCE   2  (bases 1 to 6354)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 21, 2022 from SnapGene 6.0.5
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Exported  
            2-NOV-2020 from NEBuilder https://nebuilder.neb.com"
COMMENT     NEBuilder-generated oligos (UPPERCASE = gene-specific, lowercase = 
            overlap)
            CrChR2_pAAV_fwd: 
            TAAGAATTCCGTACAAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTG 
            CrChR2_pAAV_fwd 3'Tm: 70.0  3'Ta: 72.0
            CrChR2_pAAV_rev: ctcgcccttgctcacGGCGGCCGCTGGCACGGC
            CrChR2_pAAV_rev 3'Tm: 74.4  3'Ta: 72.0
            YFP_fwd: GTGAGCAAGGGCGAGGAG
            YFP_fwd 3'Tm: 62.1  3'Ta: 60.8
            YFP_rev: gctttgtacggaattcttaCTTGTACAGCTCGTCCATG
            YFP_rev 3'Tm: 57.3  3'Ta: 60.8
FEATURES             Location/Qualifiers
     source          1..6354
                     /plasmid="New_Assembly"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    28..616
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(499..510)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    636..1112
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   1158..1298
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      1373..1828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2110..2214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2215..3075
                     /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"
     rep_origin      3246..3834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4236..4683
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      4696..4705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4702..5628
                     /codon_start=1
                     /label=CapChR1
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFDILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVICIHLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKECWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     misc_feature    4888..4890
                     /label=S63D
     misc_feature    5095..5097
                     /label=L132C
     misc_feature    5176..5178
                     /label=T159C
     misc_feature    5473..5475
                     /label=N258E
     CDS             join(5641..6354,1..3)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /label=eYFP
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
ORIGIN
        1 taagaattcc gtacaaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
       61 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
      121 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
      181 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
      241 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
      301 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
      361 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
      421 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
      481 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
      541 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
      601 tgggccgcct ccccgcatcg ataccggctc gagacgggtg gcatccctgt gacccctccc
      661 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
      721 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
      781 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
      841 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
      901 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
      961 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     1021 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     1081 aggcgtgaac cactgctccc ttccctgtcc ttctcgagct gattttgtag gtaaccacgt
     1141 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     1201 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     1261 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     1321 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     1381 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1441 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1501 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1561 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     1621 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1681 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     1741 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1801 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     1861 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1921 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1981 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     2041 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2101 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2161 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2221 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2281 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2341 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2401 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2461 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2521 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2581 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2641 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2701 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2761 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2821 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2881 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2941 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3001 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3061 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3121 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3181 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3361 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3421 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3481 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3541 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3601 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3661 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3721 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3781 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3841 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     3901 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
     3961 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     4021 ttcctgcggc cgcacgcgtt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     4081 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     4141 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg cagcaaatgg
     4201 ttaattaatc tagactgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg
     4261 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
     4321 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
     4381 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
     4441 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
     4501 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
     4561 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
     4621 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
     4681 cagtcgagag gatccgccac catggactat ggcggcgctt tgtctgccgt cggacgcgaa
     4741 cttttgttcg ttactaatcc tgtggtggtg aacgggtccg tcctggtccc tgaggatcaa
     4801 tgttactgtg ccggatggat tgaatctcgc ggcacgaacg gcgctcagac cgcgtcaaat
     4861 gtcctgcagt ggcttgcagc aggattcgac attttgctgc tgatgttcta tgcctaccaa
     4921 acctggaaat ctacatgcgg ctgggaggag atctatgtgt gcgccattga aatggttaag
     4981 gtgattctcg agttcttttt tgagtttaag aatccctcta tgctctacct tgccacagga
     5041 caccgggtgc agtggctgcg ctatgcagag tggctgctca cttgtcctgt catctgtatc
     5101 cacctgagca acctcaccgg cctgagcaac gactacagca ggagaaccat gggactcctt
     5161 gtctcagaca tcgggtgtat cgtgtggggg gctaccagcg ccatggcaac cggctatgtt
     5221 aaagtcatct tcttttgtct tggattgtgc tatggcgcga acacattttt tcacgccgcc
     5281 aaagcatata tcgagggtta tcatactgtg ccaaagggtc ggtgccgcca ggtcgtgacc
     5341 ggcatggcat ggctgttttt cgtgagctgg ggtatgttcc caattctctt cattttgggg
     5401 cccgaaggtt ttggcgtcct gagcgtctat ggctccaccg taggtcacac gattattgat
     5461 ctgatgagta aagagtgttg ggggttgttg ggacactacc tgcgcgtcct gatccacgag
     5521 cacatattga ttcacggaga tatccgcaaa accaccaaac tgaacatcgg cggaacggag
     5581 atcgaggtcg agactctcgt cgaagacgaa gccgaggccg gagccgtgcc agcggccgcc
     5641 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5701 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     5761 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     5821 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     5881 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     5941 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     6001 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     6061 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     6121 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     6181 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     6241 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     6301 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caag
//
