LOCUS       sequence-494909-        6345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This vector expresses Green Fluorescent Protein under the control of
            the Tree Shrew Cerebellin 2 Promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6345)
  TITLE     CBLN2 Promoter Enables Genetic Access to Wide Field Neurons of the 
            Tree Shrew Superior Colliculus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6345)
  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            "Unpublished"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: AAV-tsC2Pro-GFP
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1973..1997
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1998..2007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2004..2006,2007..2009,2010..2723)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2004..2006,2007..2009,2010..2720)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2049..2070)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2310..2329)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2657..2678
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2724..2748)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2758..3346
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2811..2831)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3229..3240)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3376..3393)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3382..3589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3597..3737
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3597..3726
                     /label=AAV2 ITR
     rep_origin      3812..4267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4109..4130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4284..4303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4403..4425
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4463..4481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4549..4653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4654..4722,4723..5514)
                     /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(4872..4891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5685..6273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6174..6193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6335..6345,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttggca gaggccagca ctgttttttg
      181 gggtgaaagc ccatcagcaa acctcgtcct tgtctctagg actccccctc ttctgccaag
      241 tccttgcccg cagcctgcgg cagggacgcc ccagcgcccg taggtggcgc cccgcgtccg
      301 tgcttgtgcg tctcgcccgc gctccaggcg cagctcggaa acccggcgcg tctccccggt
      361 catcagggct cagaacccgc attttaataa ataagcacca gtgccaacgc gctccgcgag
      421 cccggaggtt cagaggagca gacacataaa cccagagagg cacctcttcg atccctgccc
      481 aaagcctggc atttcccaaa ctggagattc cggaaccaca gccctcccat cattacgtcc
      541 ccaggcgttt tctctccatc tcagtctttt ccttaagaga actgtttttt ccccccttca
      601 ttatgaaagt ggtcacacta tccaagacta agacttggag ggaatttgga aaagaaaaga
      661 aacaatctaa aagaagagac gcgaccggtg cttccgaggg cgcctaatct tagcggcgtc
      721 tgggaggatt tcgctcgggg tgtttgggtg agagcctggc cagggcgttg cagggtgcgg
      781 cgggtgcggg ctggggcatc agggcgggag gagcgcggtg cgcggagtcc tcagggtgag
      841 ggcagcgctc ggcggagccc tcgggagtct ggcggggagg ctgggactga gggtccccgc
      901 cagagccgcg ctgtggacac gcgggttagg gagcactcgg gccgcgcgga gcagccggcc
      961 gaagctgggt gagggctccg tgccgggcgc gaagccttcg gtagtggcgc tgggcttctg
     1021 cggtgccggg tcgcagcgca cgggaccgga gctggggccg gagctggggc cggagccggg
     1081 gccagagccc gggccggacc gggcgcagcg gcgcacgagt gcggggcgtc ggtgcgtgcg
     1141 gagcccgctc ggccggcgcg ggaaagcggc agggcggggc ggcgcggtta ttggccgtgc
     1201 gggcagaagg gtcgcgctcc gagtcctcct cccgccagct cccgtccgcc tcattttctc
     1261 tgagcagaac ttcgcggaca gcagtgctcg cccatcatgg cagttccagg taacggagcc
     1321 cggccgtgcg cttcgggatc gtggcttcat ggctttcttg cgtacgagct aacgaggagc
     1381 gtggcgagcc tgcgttctgg ggtgcaagag ccagaggcag cgggcgcgcc tctcggtcgg
     1441 tctgccctgg gctgacgaca gggtgttagc cgagaagcaa cttctgtctt ctcggaaaga
     1501 atattgcaag agttgttcgg aggccgtggg cggggagggg ggaggcttct gtgcgttttg
     1561 tttctgggct gggaattgaa acgccctgtc tggcttagtg ggagaagagg gacggggaaa
     1621 gtcgaaacgg acgcgtaagt tagccgaacc ccacaagtgc tgcctcccag ccctggaggc
     1681 cggaccggct cagaacaata gctggggaat acgccgagcg tggattcccg tcgctggagc
     1741 ctgcccttgg cagagagggg gacgcggggt ttctcgcagg gatcagacag cggctgcgga
     1801 gacctgaaag tgggtatttg gctttggtcc ctctgtctct cgctccttcc tcagtccctc
     1861 tgtctttgct tttttttttt tttctctctc cctttttagt ctttttcacc ctaagtaact
     1921 tcctgggatt tgttttttgg gtggaaaatg gatagaatct ttactaaagg gaccaagttt
     1981 gtacaaaaaa gcaggctgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     2041 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2101 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2161 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2221 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2281 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2341 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2401 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2461 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2521 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2581 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2641 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2701 gcatggacga gctgtacaag taaacccagc tttcttgtac aaagtgggaa ttccgataat
     2761 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2821 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2881 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2941 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3001 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3061 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3121 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3181 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3241 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3301 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ggaattccta
     3361 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3421 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3481 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3541 aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag ggccgcagga
     3601 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3661 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3721 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3781 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4261 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4321 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4381 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4441 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4501 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4861 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5341 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca
     5401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6301 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcag
//
