LOCUS       Exported                6286 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 6286)
  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..6286
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     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="pause site
                     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
                     human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff"
     misc_feature    888..893
                     /label=Misc_Feature_2
                     /note="Kozak"
                     /note="Kozak Kozak;color: #a6acb3??color: #a6acb3
                     vntifkey=21"
                     /note="color: #3366cc"
     CDS             894..2243
                     /codon_start=1
                     /label=GCaMP6f
                     /label=CDS_1
                     /note="G6f"
                     /note="GCaMP6f GCaMP6f;color: #00ff00??color: #00ff00 
                     vntifkey=4"
                     /note="color: #ff9966"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2244..2603
                     /codon_start=1
                     /label=EE-RR
                     /label=GCC26
                     /note="GCC26"
                     /note="vntifkey=4"
                     /note="color: #ff9966"
                     /translation="ELGGSGGSGGTGGSGGSGGTGGSGGSGGTLEIEAAFLEQENTALE
                     TEVAELEQEVQRLENIVSQYETRYGPLGSLEIRAAFLRRRNTALRTRVAELRQRVQRLR
                     NIVSQYETRYGPL*EF"
     misc_feature    2622..3210
                     /label=WPRE
                     /note="WPRE"
                     /note="WPRE
                     woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /note="color: #a6acb3"
     CDS             complement(3093..3104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa recognition and cleavage site
                     /note="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /note="color: #993366"
                     /translation="IEGR"
     polyA_signal    3249..3456
                     /label=bGH poly(A) signal
                     /note="bGH poly(A) signal"
                     /note="bGH poly(A) signal
                     bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3613..3629)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="M13 fwd
                     common sequencing primer, one of multiple similar variants"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      3771..4226
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 ori
                     f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4558..4662
                     /gene="bla"
                     /note="bla promoter"
                     /note="AmpR promoter"
                     /note="color: #ffffff"
     CDS             4663..5523
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="bla"
                     /note="AmpR
                     confers resistance to ampicillin, carbenicillin, and 
                     related antibiotics"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5694..6282
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="ori
                     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 gagttcactc 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 ggagttctcc ctatttgaca
     1861 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc
     1921 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg
     1981 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca
     2041 cggaagaaga aattagagaa gcgttcggtg 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 gcagccttaa
     3601 ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3661 ccaacttaat cgccttgcag cacatccccc tttcgccagc ctggcgtaat agcgaagagg
     3721 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     3781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3841 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3961 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4021 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4081 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4141 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4201 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     4261 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4321 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4381 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4441 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4501 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4561 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4621 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4681 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4741 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4801 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4861 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4921 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4981 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5041 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5101 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5161 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5221 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5281 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5341 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5401 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5461 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5521 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5581 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5641 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5701 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5761 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5821 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5881 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5941 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6001 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6061 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6121 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6181 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6241 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacg
//
