LOCUS       Exported                7049 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV-mediated expression of Jaws-GFP-ER2 under the EF1-alpha promoter
            (1.1kb short version)..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1-alpha-1.1-Jaws-KGC-GFP-ER2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  AUTHORS   Chuong AS, Miri ML, Busskamp V, Matthews GA, Acker LC, Sorensen AT, 
            Young A, Klapoetke NC, Henninger MA, Kodandaramaiah SB, Ogawa M, 
            Ramanlal SB, Bandler RC, Allen BD, Forest CR, Chow BY, Han X, Lin Y,
            Tye KM, Roska B, Cardin JA, Boyden ES
  TITLE     Noninvasive optical inhibition with a red-shifted microbial 
            rhodopsin.
  JOURNAL   Nat Neurosci. 2014 Aug;17(8):1123-9. doi: 10.1038/nn.3752. Epub 2014
            Jul 6.
  PUBMED    24997763
REFERENCE   2  (bases 1 to 7049)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7049
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        314..1492
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          545..1483
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1440..1460
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2420..3133
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2462..2483)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2723..2742)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3070..3091
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3182..3770
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3235..3255)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3653..3664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3790..4266
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3989..4008)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4312..4452
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4312..4441
                     /label=AAV2 ITR
     rep_origin      4527..4982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4614..4633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4824..4845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4999..5018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5118..5140
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5178..5196)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5264..5368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5369..6229
                     /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(5587..5606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6400..6988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6889..6908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aatggttaat taaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
      361 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
      421 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      481 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      541 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
      601 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
      661 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
      721 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
      781 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
      841 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
      901 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcgacgag
      961 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     1021 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     1081 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     1141 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     1201 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     1261 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     1321 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     1381 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     1441 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggatccgc
     1501 caccatgacc gccgtgagca ccacagccac taccgtgctg caggccacac agagcgacgt
     1561 gctgcaggag atccagtcca acttcctgct gaatagctcc atctgggtga acattgctct
     1621 ggccggagtg gtcatcctgc tgtttgtggc catggggagg gatctggaat cccctagagc
     1681 taagctgatc tgggtggcca caatgctggt gccactggtg tctatttcta gttacgctgg
     1741 actggccagt gggctgactg tgggcttcct gcagatgcca cctggacacg ctctggccgg
     1801 acaggaggtg ctgagcccat ggggccggta tctgacatgg actttctcca ctcccatgat
     1861 cctgctggct ctgggactgc tggccgacac cgatattgcc agcctgttca ccgccatcac
     1921 aatggacatt ggcatgtgcg tgacaggact ggccgctgcc ctgatcacta gctcccatct
     1981 gctgcgctgg gtgttctacg gaatttcttg tgctttcttt gtggccgtgc tgtatgtgct
     2041 gctggtgcag tggccagctg atgctgaggc tgctgggacc agtgaaatct ttggcactct
     2101 gaggattctg accgtggtgc tgtggctggg gtaccctatc ctgttcgctc tgggctctga
     2161 gggagtggcc ctgctgagtg tgggagtgac cagctgggga tactccggac tggacatcct
     2221 ggctaaatac gtgttcgcct ttctgctgct gagatgggtg gctgccaatg aaggcacagt
     2281 gtctgggagt ggaatgggaa tcgggtccgg aggagctgct ccagccgacg atcgaccggt
     2341 agtaaaatcc agaattactt ctgaagggga gtatatccct ctggatcaaa tagacatcaa
     2401 tgttgcacct gcaggggcag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2461 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2521 gggcgatgcc acctacggca agctgaccct gaagttcatt tgcaccaccg gcaagctgcc
     2581 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2641 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2701 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2761 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2821 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2881 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2941 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3001 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3061 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3121 cgagctgtac aagttctgct acgagaatga agtgtaagaa ttccgtacaa agcttatcga
     3181 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3241 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3301 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3361 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3421 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3481 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3541 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3601 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3661 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3721 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3781 gctcgagacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3841 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3901 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3961 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     4021 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     4081 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     4141 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4201 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4261 gtccttctcg agctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     4321 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4381 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4441 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4501 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4561 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     4621 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4681 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4741 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4801 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     4861 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     4921 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4981 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5041 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5101 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5161 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5221 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5281 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5341 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5401 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5461 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5521 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5581 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5641 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5701 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5761 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5821 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5881 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5941 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6001 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6061 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6121 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6181 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6241 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6301 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6721 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6781 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6841 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6901 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6961 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7021 ggccttttgc tggccttttg ctcacatgt
//
