pAAV-EF1_-DIO-GRAB_NPY1.0

LOCUS       pAAV-EF1_-DIO-GRAB_NPY1.0               7775 bp DNA     circular SYN 23-JAN-2024
DEFINITION  Expresses the genetically-encoded fluorescent neuropeptide Y (NPY) 
            sensor GRAB_NPY1.0 in a cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1_-DIO-GRAB_NPY1.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7775)
  TITLE     A toolkit of highly selective and sensitive genetically encoded 
            neuropeptide sensors
REFERENCE   2  (bases 1 to 7775)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 8, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   224..353
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        454..1632
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          685..1623
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1580..1600
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1663..1696
                     /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    1747..1780
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1793..3970)
                     /codon_start=1
                     /label=NPY1.0
                     /translation="METDTLLLWVLLLWVPGSTGDTSLYKKVGTTGNSTLFSQVENHSV
                     HSNFSEKNAQLLAFENDDCHLPLAMIFTLALAYGAVIILGVSGNLALIIIILKQKEMRN
                     VTNILIVNLSFSDLLVAITCLPFTFVYTLMDHWVFGEAMCKLNPFVQCVSITVSIFSLV
                     LIAVERHQLIINPRGWRPNNRHAYVGIAVIWVLAVASSLPFLIYQVMTDEPFQNVTLDA
                     YKDKYVCFDQFPSDSHRLSYTTLLLVLQYFGPLCFIFICYFKIYIRLKRRNNMMPPSRR
                     GPDAVAAPPGGTERRPNGLGPERSAGPGGAEAEPLPTQLNGAPGEPAPAGPRDTDALDL
                     EEGGNVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSV
                     QSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVRKGEELFTGVVPILVE
                     LDGDVNGHKFSVSGEGEGDATEGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPD
                     HMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGN
                     ILGHKLEYNTGAAARWRGRQNETKRINIMLLSIVVAFAVCWLPLTIFNTVFDWNHQIIA
                     TCNHNLLFLLCHLTAMMSTCVNPIFYGFLNKNFQRDLQFFFNFCDFRSRDDDYETIAMS
                     TMHTDVSKTSLKQASPVAFKKINNNDDNEKI"
     primer_bind     2325..2344
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2584..2605
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2711..2732)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3883..3907)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(3908..3970)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     regulatory      3967..3976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3983..4016)
                     /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(4067..4100)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4125..4713
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4178..4198)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4596..4607)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4745..5221
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4944..4963)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5261..5401
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5261..5390
                     /label=AAV2 ITR
     rep_origin      5476..5931
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5563..5582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5773..5794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5948..5967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6067..6089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6127..6145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6213..6317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6318..7178
                     /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(6536..6555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7349..7775,1..162)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
       61 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
      241 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      301 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
      361 cacgcgtaag ctttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa
      421 tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag
      481 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      541 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      601 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      661 aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc
      721 tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt
      781 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
      841 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
      901 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
      961 ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     1021 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     1081 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     1141 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     1201 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     1261 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     1321 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     1381 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     1441 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     1501 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     1561 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     1621 caggtgtcgt gaggtaccgg atcctctaga gtcgactccg gaataacttc gtataggata
     1681 ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac
     1741 ctcttaataa cttcgtatag catacattat acgaagttat ggcgcatatg tcttagattt
     1801 tttcattatc atcattgttg ttgatttttt taaatgcgac tgggcttgct tgcttcaaag
     1861 aagttttgga aacatctgtg tgcatcgtgg acatggctat tgtttcataa tcatcatccc
     1921 gagaccggaa atcacaaaag ttgaagaaga actgcaagtc tctctggaag tttttgttca
     1981 ggaacccata aaatatgggg ttgacacaag tggacatcat tgctgtgagg tggcagagca
     2041 ggaataacag attgtggttg caggtagcaa tgatctgatg attccaatca aacacagtgt
     2101 taaagatggt aagagggagc cagcagactg caaatgctac cacaatggag agcagcatga
     2161 tattgattct tttggtttcg ttctgccgcc cgcgccagcg tgctgctgct ccggtgttgt
     2221 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     2281 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     2341 cgtccttgaa gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga
     2401 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     2461 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     2521 tcagcttgcc ctcggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt
     2581 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     2641 ttctcaccat gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc
     2701 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg
     2761 aaagtttgga ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2821 tgggggtgtt ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc
     2881 ggatgtggaa gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgtttc
     2941 ctccctcctc caggtccagc gcgtcggtgt cgcgcggccc ggccggcgcg ggctcgccag
     3001 gggcgccgtt gagctgggtg ggcagcggtt cggcctctgc gccccccggg cccgcgctgc
     3061 gctcggggcc cagaccgttg ggcctgcgct cggtgccccc cggcggcgcg gcgacggcgt
     3121 ccggaccccg gcggctgggt ggcatcatgt tgtttctcct ttttaggcgt atatatatct
     3181 tgaagtagca aataaatata aaacaaagtg gaccaaaata ctgcagcacc aagaggagag
     3241 tggtataaga caacctatga gagtccgatg gaaattgatc aaagcacacg tatttgtctt
     3301 tgtacgcatc aagtgttaca ttttggaacg gctcatcagt cattacttgg tagatcagga
     3361 aaggcaaaga agaagccaca gcaaggaccc aaatcacagc aatacctaca taagcatgtc
     3421 tattatttgg tctccaccct cgagggttga ttatcagctg atgtcgttcc acagcaatga
     3481 gaaccagaga gaaaatggac acagtgattg aaacacattg cacaaaagga ttcaacttac
     3541 acatcgcctc accgaagacc cagtggtcca ttaatgtgta gacaaatgta aaggggagac
     3601 acgtgatggc aacaagcaag tctgagaagg aaaggttcac aatcaggatg ttggtaacat
     3661 ttctcatctc cttttgtttc aagatgatta tgatcaaggc caggtttcca gagacaccaa
     3721 gaatgatcac agctccataa gcaagagcta aggtaaatat catggccaag ggcagatgac
     3781 aatcatcatt ttcaaaagcc agaagctggg cattcttctc tgagaaatta gagtggactg
     3841 aatgattttc aacctgggaa aataatgttg aattaccggt ggtgccaact tttttgtaca
     3901 aacttgtgtc accagtggaa cctggaaccc agagcagcag tacccatagc aggagtgtgt
     3961 ctgtctccat ggtggcgcta gcataacttc gtataaagta tcctatacga agttatttgc
     4021 cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa
     4081 tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa cctctggatt
     4141 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4201 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4261 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4321 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4381 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4441 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4501 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     4561 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4621 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4681 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     4741 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     4801 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     4861 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     4921 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4981 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     5041 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     5101 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     5161 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     5221 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     5281 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     5341 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     5401 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     5461 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5521 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     5581 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5641 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5701 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5761 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     5821 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     5881 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5941 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     6001 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6061 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     6121 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6181 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6241 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6301 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6361 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6421 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6481 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6541 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6601 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6661 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6721 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6781 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6841 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6901 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6961 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7021 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7081 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7141 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7201 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7261 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7321 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7381 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7441 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7501 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7561 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7621 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7681 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7741 tatgagaaag cgccacgctt cccgaaggga gaaag
//