LOCUS       sequence-76825-e        6390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer vector with mammalian codon-optimized ChD-citrine 
            between AAV2 ITR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6390)
  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 6390)
  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-oChD-citrine
FEATURES             Location/Qualifiers
     source          1..6390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(264..269,270..287,288..294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   380..509
                     /label=AAV2 ITR
     promoter        591..1037
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(2146..2148,2149..2151,2152..2862)
                     /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(2191..2212)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2452..2471)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2799..2820
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2899..3487
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2952..2972)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3370..3381)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3490..3506)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3491..3507)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3536..3657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3547..3566)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3601..3620
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3819..3948
                     /label=AAV2 ITR
     primer_bind     complement(3967..3984)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3967..3983)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3976..3998)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4125..4580
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4212..4231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4422..4443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4606..4710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4711..4779,4780..5571)
                     /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(4929..4948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5742..6330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6231..6250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg
      601 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccacccac
      661 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      721 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      781 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      841 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      901 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      961 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
     1021 cgtgcctgag agcgcagtcg aattcaagct gctagcaagg atccaccggt gaccatggtg
     1081 agcagaagac cctggctgct ggccctggcc ctggccgtgg ccctggccgc cggcagcgcc
     1141 ggcgccagca ccggcagcga cgccaccgtg cccgtggcca cccaggacgg ccccgactac
     1201 gtgttccaca gagcccacga gagaatgctg ttccagacca gctacaccct ggagaacaac
     1261 ggcagcgtga tctgcatccc caacaacggc cagtgcttct gcctggcctg gctgaagagt
     1321 aacggcacca acgccgagaa gctggccgcc aacatcctgc agtggatcac cttcgccctg
     1381 agcgccctgt gcctgatgtt ctacggctac cagacctgga agagtacctg cggctgggag
     1441 gagatctacg tggccaccat cgagatgatc aagttcatca tagagtactt ccacgagttc
     1501 gacgagcccg ccgtgatcta cagcagcaac ggcaacaaga ccgtgtggct gagatacgcc
     1561 gagtggctgc tgacctgccc cgtgatcctg atccacctga gcaacctgac cggcctggcc
     1621 aacgactaca acaagagaac catgggcctg ctggtgagcg acatcggcac catcgtgtgg
     1681 ggggctacca gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg
     1741 tgctatggcg cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact
     1801 gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc
     1861 tggggtatgt tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc
     1921 tatggctcca ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg
     1981 ttgggacact acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc
     2041 aaaaccacca aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac
     2101 gaagccgagg ccggagccgt ggcaccgtcg ccaccattgc tcgagatggt gagcaagggc
     2161 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2221 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2281 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     2341 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2401 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2461 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2521 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2581 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2641 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2701 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     2761 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2821 accgccgccg ggatcactct cggcatggac gagctgtaca agtaggctag ccccgaattc
     2881 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2941 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3001 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3061 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3121 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3181 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3241 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3301 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3361 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3421 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3481 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga
     3541 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3601 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3661 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3721 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     3781 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     3841 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3901 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc
     3961 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4021 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4081 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4201 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4321 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4441 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4501 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4561 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4681 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4741 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4801 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4861 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4921 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4981 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5041 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5101 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5161 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5221 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5281 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5341 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5401 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5461 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5521 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5581 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5641 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5881 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6001 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6121 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6361 ctggcctttt gctggccttt tgctcacatg
//
