LOCUS       Exported                6218 bp ds-DNA     circular SYN 30-JUN-2020
DEFINITION  Ligation of KA2(1-150)-12-ChR90-3xFLAG-2A-GCaMPV2 into 
            pAAV.hSynapsin.E BamHI/HindIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 22, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     SECID/File created by SciEd Central, Scientific
FEATURES             Location/Qualifiers
     source          1..6218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          58..187
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          882..2615
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /note="color: #ffffff"
     source          894..2243
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          894..2243
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          894..2243
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          3465..3594
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          3595..3628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     repeat_region   58..187
                     /label=AAV2 ITR (alternate)
                     /note="AAV2 ITR (alternate)"
                     /note="Functional equivalent of wild-type AAV2 ITR"
                     /note="color: #ffcc99"
     polyA_signal    243..291
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    305..396
                     /label=pause site
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
                     /note="color: #a6acb3"
     promoter        432..879
                     /label=Syn promoter
                     /note="hSyn promoter"
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             894..2243
                     /codon_start=1
                     /label=GcaMP7f
                     /note="GcaMP7f"
                     /note="color: #00ff00"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2244..2594
                     /codon_start=1
                     /label=EE-RR
                     /label=GCC26
                     /note="GCC26"
                     /note="vntifkey=4"
                     /note="color: #ff9966"
                     /note="color: #993366"
                     /translation="ELGGSGGSGGTGGSGGSGGTGGSGGSGGTLEIEAAFLEQENTALE
                     TEVAELEQEVQRLENIVSQYETRYGPLGSLEIRAAFLRRRNTALRTRVAELRQRVQRLR
                     NIVSQYETRYGPL"
     misc_feature    2622..3210
                     /label=WPRE
                     /note="WPRE"
                     /note="woodchuck hepatitis virus?posttranscriptional
                     regulatory element"
                     /note="color: #ffffff"
     polyA_signal    3249..3456
                     /label=bGH poly(A) signal
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3465..3594
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
                     /note="color: #ffcc99"
     rep_origin      3703..4158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4490..4594
                     /gene="bla"
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4595..5455
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments: 
                     ???1:?4595?..?4663?/?#ccffcc?/?signal?sequence 
                     ???2:?4664?..?5455?/?#ccffcc
                     Cleavage site after base 4663"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5626..6214
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
       61 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
      121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      181 ggttcctgcg gccgcacgcg ttctagaaat attaaggtac gggaggtact tggagcggcc
      241 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      301 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      361 tccccagtgc aagtgcaggt gccagaacat ttctctattc tagactcgag ctgcagaggg
      421 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      481 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      541 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      601 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      661 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      721 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      781 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      841 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cggatccgcc accatgggtt
      901 ctcatcatca tcatcatcat ggtatggctt ccatgactgg tggacagcaa atgggtcggg
      961 atctgtacga cgatgacgat aaggatctcg ccaccatggt cgactcatca cgtcgtaagt
     1021 ggaataagac aggtcacgca gtcagagcta taggtcggct gagttcactg gagaacgtct
     1081 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
     1141 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg
     1201 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc
     1261 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1321 tcggcatgga cgagctgtac aagggcggaa ccggagggag catggtgagc aagggcgagg
     1381 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1441 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     1501 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1561 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1621 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact
     1681 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1741 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctgc
     1801 cggaccaact gactgaagag cagatcgcag aatttaaaga gctgttctcc ctatttgaca
     1861 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc
     1921 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg
     1981 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa tacacagaca
     2041 gtgaagaaga aattagagaa gcgttccgtg tgtttgataa ggatggcaat ggctacatca
     2101 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     2161 ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac tacgaagagt
     2221 ttgtacaaat gatgacagcg aaggagctcg ggggcagcgg tgggtctgga gggactgggg
     2281 gtagcggtgg tagcggtggg acagggggaa gcggcggtag cggtgggaca ctggaaatcg
     2341 aggccgcttt cctggagcag gagaacaccg ccctggagac agaggtggct gagctggagc
     2401 aggaggtgca gcggctggag aacatcgtgt ctcagtacga gaccagatac ggaccactgg
     2461 gatccctgga gatccgggcc gcttttctga ggagacggaa caccgccctg aggacaaggg
     2521 tggctgagct gaggcagaga gtgcagcggc tgcgcaacat cgtgagccag tacgagacac
     2581 gctacggacc tctgtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca
     2641 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2701 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2761 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2821 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2881 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2941 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3001 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     3061 ttctgcgcgg gacgcccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3121 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3181 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgactagag ctcgctgatc
     3241 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3301 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3361 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3421 ggaggattgg gaagacaata gcaggcatgc tggggagaga tctgaggaac ccctagtgat
     3481 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3541 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg
     3601 agtggccccc cccccccccc cccccctgca gcctggcgta atagcgaaga ggcccgcacc
     3661 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     3721 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3781 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3841 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3901 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3961 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4021 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4081 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4141 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4201 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4261 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4321 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4381 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4441 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4501 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4561 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4621 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4681 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4741 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4801 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4861 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4921 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4981 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5041 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5101 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5161 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5221 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5281 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5341 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5401 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5461 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5521 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5581 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5641 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5701 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5761 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5821 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5881 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5941 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6001 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6061 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6121 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6181 gtgatgctcg tcaggggggc ggagcctatg gaaaaacg
//
