LOCUS       sequence-211134-        6034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ChR2-YFP reporter vector of iTango and Cal-Light system for AAV 
            production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  TITLE     fluorescent protein reporter vectors of iTango and Cal-Light system 
            for AAV production
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6034)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-TRE-ChR2-YFP
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    join(298..316,317..339,340..358,359..381,382..400,401..423,
                     424..442,443..465,466..484,485..507,508..526,527..549,
                     550..568)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        601..639
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             760..1686
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             join(1699..1701,1702..2412)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1741..1762)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2002..2021)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2349..2370
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2449..3037
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2502..2522)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2920..2931)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3072..3089)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3078..3285
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3308..3437
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3499..4087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3579..3598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4258..5049,5050..5118))
                     /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     4881..4900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5119..5223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5291..5309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5347..5369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5469..5488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5505..5960)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5642..5663)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5854..5873
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga attcggggcc
      181 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      241 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc
      301 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      361 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      421 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      481 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      541 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga
      601 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
      661 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      721 ctccgcggcc ccgaattcga gctcggtacc cggggatcca tggactatgg cggcgctttg
      781 tctgccgtcg gacgcgaact tttgttcgtt actaatcctg tggtggtgaa cgggtccgtc
      841 ctggtccctg aggatcaatg ttactgtgcc ggatggattg aatctcgcgg cacgaacggc
      901 gctcagaccg cgtcaaatgt cctgcagtgg cttgcagcag gattcagcat tttgctgctg
      961 atgttctatg cctaccaaac ctggaaatct acatgcggct gggaggagat ctatgtgtgc
     1021 gccattgaaa tggttaaggt gattctcgag ttcttttttg agtttaagaa tccctctatg
     1081 ctctaccttg ccacaggaca ccgggtgcag tggctgcgct atgcagagtg gctgctcact
     1141 tgtcctgtca tccttatccg cctgagcaac ctcaccggcc tgagcaacga ctacagcagg
     1201 agaaccatgg gactccttgt ctcagacatc gggactatcg tgtggggggc taccagcgcc
     1261 atggcaaccg gctatgttaa agtcatcttc ttttgtcttg gattgtgcta tggcgcgaac
     1321 acattttttc acgccgccaa agcatatatc gagggttatc atactgtgcc aaagggtcgg
     1381 tgccgccagg tcgtgaccgg catggcatgg ctgtttttcg tgagctgggg tatgttccca
     1441 attctcttca ttttggggcc cgaaggtttt ggcgtcctga gcgtctatgg ctccaccgta
     1501 ggtcacacga ttattgatct gatgagtaaa aattgttggg ggttgttggg acactacctg
     1561 cgcgtcctga tccacgagca catattgatt cacggagata tccgcaaaac caccaaactg
     1621 aacatcggcg gaacggagat cgaggtcgag actctcgtcg aagacgaagc cgaggccgga
     1681 gccgtgccag cggccgccgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1741 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1801 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1861 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
     1921 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1981 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2041 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2101 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2161 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2221 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2281 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     2341 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2401 gagctgtaca agttctgcta cgagaacgag gtgtaaaagc ttatcgataa tcaacctctg
     2461 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2521 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2581 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2641 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2701 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2761 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2821 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2881 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2941 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3001 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3061 agatcgatct gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3121 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3181 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3241 cagcaagggg gaggattggg aagacaatag caggcatgct ggggacacgt gcggaccgag
     3301 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3361 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     3421 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3901 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4141 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4201 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4261 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4321 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4381 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4441 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4501 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4561 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4621 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4681 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4741 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4801 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4861 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4921 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4981 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5041 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5101 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5161 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5221 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5281 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5341 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5401 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5461 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
     5521 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
     5581 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
     5641 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5701 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     5761 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5821 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5881 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5941 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     6001 agatgcgtaa ggagaaaata ccgcatcagg cgcc
//
