pAAV-hSyn-DIO-GRAB_UCN1.0

LOCUS       pAAV-hSyn-DIO-GRAB_UCN1.0           6866 bp DNA     circular SYN 23-JAN-2024
DEFINITION  Expresses the genetically-encoded fluorescent urocortin (UCN) sensor
            GRAB_UCN1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-GRAB_UCN1.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6866)
  TITLE     A toolkit of highly selective and sensitive genetically encoded 
            neuropeptide sensors
REFERENCE   2  (bases 1 to 6866)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 9, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        131..574
                     /label=hSyn
     protein_bind    605..638
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    689..722
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(733..2838)
                     /codon_start=1
                     /label=UCN1.0
                     /translation="METDTLLLWVLLLWVPGSTGDTSLYKKVGTTGDAALLLSLLEANC
                     SLALAEELLLDGWGVPPDPEGPYTYCNTTLDQIGTCWPQSAPGALVERPCPEYFNGIKY
                     NTTRNAYRECLENGTWASRVNYSHCEPILDDKQRKYDLHYRIALIVNYLGHCVSVVALV
                     AAFLLFLVLRSIRCLRNVIHWNLITTFILRNIAWFLLQLIDHEVHEGNEVWCRCITTIF
                     NYFVVTNFFWMFVEGCYLHTAIVMTYSTEHLRKWLFLFIGWCIPCPIIIAWAVGKLYYE
                     NEQCWFGKEAGDLVDYIYQGPVMLVLLINFVFLFNIVRILMTKLRAYTGPDTDALDLEE
                     GGNVYIKADKQKNGIKANFTIRHNCEDGGVQLATHYQQNTPIGDGPVLLPDNHYLSVQS
                     KLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVRKGEELFTGVVPILVELD
                     GDVNGHKFSVSGEGEGDATEGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHM
                     KQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNIL
                     GHKLEYNTGAAARWRGRQMRKAVKATLVLLPLLGITYMLFFVNPGEDDLSQIVFIYFNS
                     FLQSFQGFFVSVFYCFFNGEVRAALRKRWHRWQDHHALRVPVARAMSIPTSPTRISFHS
                     IKQTAAV"
     primer_bind     1199..1218
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1458..1479
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1585..1606)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2751..2775)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(2776..2838)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     regulatory      2835..2844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2851..2884)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(2935..2968)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2993..3581
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3046..3066)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3464..3475)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3613..4089
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3812..3831)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4129..4269
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4129..4258
                     /label=AAV2 ITR
     rep_origin      4344..4799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4431..4450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4641..4662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4816..4835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4935..4957
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4995..5013)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5081..5185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5186..6046
                     /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(5404..5423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6217..6805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6706..6725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccacctcta gagtcgactc
      601 cggaataact tcgtatagga tactttatac gaagttatgc agaatggtag ctggattgta
      661 gctgctatta gcaatatgaa acctcttaat aacttcgtat agcatacatt atacgaagtt
      721 atggcgcata tgtcacacag cagctgtctg cttgatgctg tggaagctga tcctggtggg
      781 cgacgtaggg atggacatgg cccgggccac aggcacccgg agggcgtggt ggtcctgcca
      841 gcggtgccac cgctttctca gggccgcgcg cacctctcca ttgaagaagc agtagaaaac
      901 ggacacaaag aaaccctgga aggactgcag gaaagagttg aagtagatga acacaatctg
      961 ggacaggtcg tcctcgccag gattgacaaa gaagagcatg taggtgatgc ccaacagggg
     1021 gaggaggacc agcgtggcct tcactgcctt cctcatctgc ctcccgcgcc agcgtgctgc
     1081 tgctccggtg ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     1141 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     1201 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggatgtagc cttcgggcat
     1261 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     1321 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     1381 gatgaacttc agggtcagct tgccctcggt ggcatcgccc tcaccctcgc cggacacgct
     1441 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     1501 cagctcctcg ccctttctca ccatgctccc tccggtaccg cccttgtaca gctcgtccat
     1561 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     1621 gttggggtct ttcgaaagtt tggactgcac gctcaggtag tggttgtcgg gcagcagcac
     1681 ggggccgtcg ccgatggggg tgttctgctg gtagtgggtg gcgagctgca cgccgccgtc
     1741 ctcgcagttg tggcggatgg tgaagttcgc cttgatgccg ttcttctgct tgtcggcctt
     1801 gatatagacg tttcctccct cctccaggtc cagcgcgtcg gtgtcggggc cggtgtatgc
     1861 tcgtaacttc gtcatcagga tcctgacgat gttaaacaga aatacaaaat tgatcaacag
     1921 cacaagcatg acggggccct ggtagatgta gtccaccaaa tcaccagctt ccttgccaaa
     1981 ccagcactgc tcattctcat agtagagttt gccaactgcc caggcgatga tgatagggca
     2041 gggaatgcac catccaatga agaggaaaag ccacttgcgc aggtgctctg tggagtacgt
     2101 catgacaatg gccgtgtgca ggtagcagcc ctccacaaac atccagaaga agttggtgac
     2161 cacaaaatag ttgaagatgg tggtgatgca gcggcaccag acctcattgc cctcgtgcac
     2221 ttcgtggtcg atgagttgca gcaggaacca cgcgatgttt ctcagaatga aggtggtgat
     2281 gaggttccag tggatcacat tcctcaggca gcggatactc cgcagcacta ggaaaagcag
     2341 gaaagcggcc accagggcca ccacggaaac acagtgaccc aggtagttga caatgagggc
     2401 gattcggtaa tgcaggtcat actttctctg cttgtcatcc aaaatgggtt cgcagtgtga
     2461 gtagttgacc cttgaggccc aggtcccgtt ctccaggcac tctctgtagg cattccgggt
     2521 cgtgttgtac ttgatgccat tgaagtactc ggggcacggt ctctctacta gggctccggg
     2581 tgcgctctgt ggccagcagg tcccgatctg gtccaaggtc gtgttgcagt aggtgtaggg
     2641 accttcgggg tccgggggca ctccccagcc gtccaggagc agctcttcgg ccagcgccag
     2701 gctgcagttg gcctccagca ggctgaggag cagcgccgcg tcaccggtgg tgccaacttt
     2761 tttgtacaaa cttgtgtcac cagtggaacc tggaacccag agcagcagta cccatagcag
     2821 gagtgtgtct gtctccatgg tggcgctagc ataacttcgt ataaagtatc ctatacgaag
     2881 ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca aagaataact
     2941 tcgtataatg tatgctatac gaagttatga attcgatatc aagcttatcg ataatcaacc
     3001 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3061 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3121 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3181 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3241 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3301 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3361 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3421 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3481 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3541 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3601 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3661 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3721 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3781 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3841 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3901 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3961 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     4021 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4081 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4141 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4201 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4261 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4321 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4381 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4441 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4501 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4561 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4621 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4681 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4741 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4801 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4861 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4921 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4981 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5041 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5101 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5161 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5221 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5281 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5341 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5401 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5461 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5521 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5581 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5641 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5701 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5761 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5821 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5881 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5941 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6001 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6061 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6121 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6841 cttttgctgg ccttttgctc acatgt
//