LOCUS       Exported                6056 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV-mediated expression of Archon1-KGC-GFP-ER2 under the Syn 
            promoter. Using bGHpA signal..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Syn-Archon1-KGC-GFP-ER2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 6056)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6056
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        360..806
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      819..828
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1692..2405
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1734..1755)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1995..2014)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2342..2363
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2454..3042
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2507..2527)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2925..2936)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3060..3267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3319..3459
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3319..3448
                     /label=AAV2 ITR
     rep_origin      3534..3989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3621..3640)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3831..3852
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4006..4025)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4125..4147
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4185..4203)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4271..4375
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4376..5236
                     /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(4594..4613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5407..5995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5896..5915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttgcaa agatggataa agttttaaac
      181 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
      241 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
      301 ggtcggcagc aaattaatta atggttaatt aatctagact gcagagggcc ctgcgtatga
      361 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      421 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      481 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      541 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      601 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      661 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg cacgggcgcg
      721 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      781 gagtcgtgtc gtgcctgaga gcgcagtcga gaggatccgc caccatggtt tctatcgcac
      841 tgcaggctgg ttatgacctc ctcggagatg gtagacctga aagtctgtgg ctcggaattg
      901 gtacactgct gatgctgatc ggaaccttct acttcctggt tagagcgtgg ggagagactg
      961 acaaggatgc acgggagtac tacgcagtga caatcctggt cagcggtatt gccagtgcag
     1021 cttatctgtc aatgttcttt ggtattggcc tcacagaggt gccagtaggt ggagagatgc
     1081 tgaacatcta ctacgcaaga tacgcacagt ggctgttcac tacaccactg ttgctgctgc
     1141 acctggcact cctggccaaa gttgatagag tgaccatcgg cactctcgtt ggcgtcgatg
     1201 ctctgatgat tgtcacaggc ctgataggaa cactgtcaca tactgccatt gccagatact
     1261 cttggtggct gttctcaact atctgtatga tcgtcgttct gtatgtcctg gccacttctc
     1321 tgcggtcagc cgctaaggag agaggcccag aggttgcctc aaccttcaac atcctcaccg
     1381 cactggttct ggtcctgtgg actgcttacc ctatcatctg gattatcggc accgagggtg
     1441 ccggagttgt gggcctggga atcgaaactt tgctgttcat ggtgctggac gtcaccgcca
     1501 aggtgggctt cggttttatc ctgctcaggt caagagccat tctgcaagat acagaagcac
     1561 ctgaaccttc agctggagct gataccggtg ccgccgaccg cccggtagta gcagtgagca
     1621 aggcggccgc caagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg
     1681 acatcaacgt ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1741 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1801 ccacctacgg caagctgacc ctgaagttca tttgcaccac cggcaagctg cccgtgccct
     1861 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1921 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1981 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2041 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2101 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2161 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2221 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2281 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2341 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2401 acaagttctg ctacgagaac gaggtgtaag aattccgtac aaagcttatc gataatcaac
     2461 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2521 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2581 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2641 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2701 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2761 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2821 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     2881 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2941 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3001 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cggctcgagc
     3061 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3121 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3181 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3241 ggaagacaat agcaggcatg ctggggatgc ggtctcgagc tgattttgta ggtaaccacg
     3301 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3361 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3421 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3481 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3541 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3601 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3661 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3721 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3781 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3841 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     3901 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3961 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4021 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4081 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4141 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4201 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4321 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4561 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4681 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4921 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4981 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5221 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5521 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
