LOCUS       sequence-444968-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer vector with mammalian codon-optimized ChIEF-citrine 
            between AAV2 ITR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  AUTHORS   Lin JY, Lin MZ, Steinbach P, Tsien RY
  TITLE     Characterization of engineered channelrhodopsin variants with 
            improved properties and kinetics.
  JOURNAL   Biophys J. 2009 Mar 4;96(5):1803-14. doi: 10.1016/j.bpj.2008.11.034.
  PUBMED    19254539
REFERENCE   2  (bases 1 to 5942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bpj.2008.11"; journalName: "Biophys J"; date: "2009-03-4-
            4"; volume: "96"; issue: "5"; pages: "1803-14"
COMMENT     Sequence Label: AAV-oChIEF-citrine
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        212..659
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1780..1782,1783..1785,1786..2496)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1825..1846)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2086..2105)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2433..2454
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2533..3121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2586..2606)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3004..3015)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3124..3140)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3125..3141)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3170..3291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3181..3200)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3235..3254
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3666..4254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3746..3765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4425..5216,5217..5285))
                     /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     5048..5067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5286..5390)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5416..5871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5553..5574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5765..5784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga
      241 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      301 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      361 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
      421 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      481 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      541 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      601 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
      661 cgaattcaag ctgctagcaa ggatccaccg gtgaccatgg tgagcagaag accctggctg
      721 ctggccctgg ccctggccgt ggccctggcc gccggcagcg ccggcgccag caccggcagc
      781 gacgccaccg tgcccgtggc cacccaggac ggccccgact acgtgttcca cagagcccac
      841 gagagaatgc tgttccagac cagctacacc ctggagaaca acggcagcgt gatctgcatc
      901 cccaacaacg gccagtgctt ctgcctggcc tggctgaaga gtaacggcac caacgccgag
      961 aagctggccg ccaacatcct gcagtggatc accttcgccc tgagcgccct gtgcctgatg
     1021 ttctacggct accagacctg gaagagtacc tgcggctggg aggagatcta cgtggccacc
     1081 atcgagatga tcaagttcat catagagtac ttccacgagt tcgacgagcc cgccgtgatc
     1141 tacagcagca acggcaacaa gaccgtgtgg ctgagatacg ccgagtggct gctgacctgc
     1201 cccgtggtcc tgatccacct gagcaacctg accggcctgg ccaacgacta caacaagaga
     1261 accatgggcc tgctggtgag cgacatcggc accatcgtgt ggggcaccac cgccgccctg
     1321 agcaagggct acgtgagagt gatcttcttc ctgatgggcc tgtgctacgg catctacacc
     1381 ttcttcaacg ccgccaaggt gtacatcgag gcctaccaca ccgtgcccaa gggcagatgc
     1441 agacaggtgg tgaccggcat ggcctggctg ttcttcgtga gctggggcat gttccccatc
     1501 ctgttcatcc tgggccccga gggcttcggc gtgctgagcg tgtacggcag caccgtgggc
     1561 cacaccatca tcgacctgat gagcaagaac tgctggggcc tgctgggcca ctacctgaga
     1621 gtgctgatcc acgagcacat cctgatccac ggcgacatca gaaagaccac caagctgaac
     1681 atcggcggca ccgagatcga ggtggagacc ctggtggagg acgaggccga ggccggcgcc
     1741 gtgaacaagg gcaccggcaa gtacgagagc agcctcgaga tggtgagcaa gggcgaggag
     1801 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1861 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1921 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     1981 ggcctgatgt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2041 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2101 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2161 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2221 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2281 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2341 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2401 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2461 gccgggatca ctctcggcat ggacgagctg tacaagtagg ctagccccga attcgatatc
     2521 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2581 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2641 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2701 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2761 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2821 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2881 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2941 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3001 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3061 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3121 catcgatacc gtcgacctcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
     3181 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3241 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3301 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3361 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     3421 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     3481 tccctctctg cgcgctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4381 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4441 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4501 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4561 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4621 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4681 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4741 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4801 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4861 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4921 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4981 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5041 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5101 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5161 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5221 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5281 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5341 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5401 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5461 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5521 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5581 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5641 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5701 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5761 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5821 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     5881 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5941 ag
//
