LOCUS       sequence-494910-        6775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This vector co-expresses Red Fluorescent Protein mScarlet3 and Cre 
            Recombinase under the control of the truncated Tree Shrew Cerebellin
            2 Promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6775)
  TITLE     CBLN2 Promoter Enables Genetic Access to Wide Field Neurons of the 
            Tree Shrew Superior Colliculus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6775)
  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            "Unpublished"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-tsC2ProT-mScarlet3-Cre
FEATURES             Location/Qualifiers
     source          1..6775
                     /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    1314..1338
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1339..1348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1345..2031
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     1743..1762
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2041..2097
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2104..2124
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2125..3153
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     protein_bind    complement(3154..3178)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3188..3776
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3241..3261)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3659..3670)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3806..3823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3812..4019
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4027..4167
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4027..4156
                     /label=AAV2 ITR
     rep_origin      4242..4697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4329..4348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4539..4560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4714..4733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4833..4855
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4893..4911)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4979..5083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5084..5152,5153..5944)
                     /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(5302..5321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6115..6703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6604..6623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6765..6775,1..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 taaccaagtt
     1321 tgtacaaaaa agcaggctgc caccatggat agcaccgagg cagtgatcaa ggagttcatg
     1381 cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     1441 ggcgagggcc gcccctacga gggcacccag accgccaagc tgagggtgac caagggtggc
     1501 cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc cagggccttc
     1561 acgaagcacc ccgccgacat ccccgactac tggaagcagt ccttccccga gggcttcaag
     1621 tgggagcgcg tgatgaactt cgaggacggc ggcgccgtgt ccgtggccca ggacacctcc
     1681 ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt ccctcctgac
     1741 ggccccgtaa tgcagaagaa gacaatgggc tgggaagcat ccaccgagcg gttgtacccc
     1801 gaggacgtcg tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc
     1861 tacctggcgg acttcaagac cacctacagg gccaagaagc ccgtgcagat gcccggcgcc
     1921 ttcaacatcg accgcaagtt ggacatcaca tcccacaacg aggactacac cgtggtggaa
     1981 cagtacgaac gctccgtggc ccgccactcc accggcggct ccggtggctc cggaagcgga
     2041 gccacgaact tctctctgtt aaagcaagca ggagatgttg aagaaaaccc cgggcctatg
     2101 ggcccaaaga agaagagaaa ggtttcgaat ttactgaccg tacaccaaaa tttgcctgca
     2161 ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat
     2221 cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg
     2281 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
     2341 tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc
     2401 cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt
     2461 tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag
     2521 gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat
     2581 cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt
     2641 atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg
     2701 ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt
     2761 agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat
     2821 ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc
     2881 agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac
     2941 ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc
     3001 ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt
     3061 ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg
     3121 gcaatggtgc gcctgctgga agatggcgat tagacccagc tttcttgtac aaagtgggaa
     3181 ttccgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3241 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3301 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3361 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3421 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3481 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3541 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3601 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3661 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3721 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3781 ggaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3841 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3901 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3961 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
     4021 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4081 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4141 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4201 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4261 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4321 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4381 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4441 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4501 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4561 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4621 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4681 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4741 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4801 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4861 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4921 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4981 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5041 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5101 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5161 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5221 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5281 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5341 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5401 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5461 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5521 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5581 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5641 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5701 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5761 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     5821 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5881 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5941 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6001 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6061 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6241 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6721 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcag
//
