LOCUS       sequence-358494-        6611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the neurotensin sensor GRAB_NTS1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  TITLE     Yulong Li lab sensors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6611)
  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            "Unpublished"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-hSyn-GRAB_NTS1.0
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1780..1801
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1907..1928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2168..2187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2738..3326
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2791..2811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3209..3220)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3358..3834
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3557..3576)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3874..4014
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3874..4003
                     /label=AAV2 ITR
     rep_origin      4089..4544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4176..4195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4386..4407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4561..4580)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4680..4702
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4740..4758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4826..4930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4931..4999,5000..5791)
                     /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(5149..5168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5962..6550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6451..6470
                     /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 gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccaccg gtcgcctcaa cagctccgcg ccgggaaccc cgggcacgcc
      721 ggccgccgac cccttccagc gggcgcaggc cggactggag gaggcgctgc tggccccggg
      781 cttcggcaac gcttcgggca acgcgtcgga gcgcgtcctg gcggcaccca gcagcgagct
      841 ggacgtgaac accgacatct actccaaggt gctggtgacc gccgtgtacc tggcgctctt
      901 cgtggtgggc acggtgggca acacggtgac ggcgttcacg ctggcgcgga agaagtcgct
      961 gcagagcctg cagagcacgg tgcattacca cctgggcagc ctggcgctgt ccgacctgct
     1021 caccctgctg ctggccatgc ccgtggagct gtacaacttc atctgggtgc accacccctg
     1081 ggccttcggc gacgccggct gccgcggcta ctacttcctg cgcgacgcct gcacctacgc
     1141 cacggccctc aacgtggcca gcctgagtgt ggagcgctac ctggccatct gccacccctt
     1201 caaggccaag accctcatgt cccgaagccg caccaagaag ttcatcagcg ccatctggct
     1261 cgcctcggcc ctgctggcgg tgcctatgct gttcaccatg ggcgagcaga accgcagcgc
     1321 cgacggccag cacgccggcg gcctggtgtg cacccccacc atccacactg ccaccgtcaa
     1381 ggtcgtcata caggtcaaca ccttcatgtc cttcatattc cccatggtgg tcatctcggt
     1441 cctgaacacc atcatcgcca acaagctgac cgtcatggta cgccaggcgg ccgagcagct
     1501 caacggcgcc cctggcgagc ccgcgccggc cgggccgcgc gacaccgacg cgctggacct
     1561 ggaggaggga ggaaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcggc
     1621 gttcaatatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca
     1681 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca
     1741 gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1801 gaccgccgcc gggatcactt ggggcatgga cgagctgtac aagggcggta ccggagggag
     1861 catggtgaga aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1921 cggcgacgta aacggccaca agttcagcgt gggcggcgag ggtgagggcg atgccaccga
     1981 gggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2041 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2101 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt
     2161 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2221 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2281 caagctggag tacaacaccg gagcagcagc acgctggcgc gggcgggagc ctaatagggt
     2341 ccaggccctg cggcacggcg tgcgcgtcct acgtgcagtg gtcatcgcct ttgtggtctg
     2401 ctggctgccc taccacgtgc ggcgcctcat gttctgctac atctcggatg agcagtggac
     2461 tccgttcctc tatgacttct accactactt ctacatggtg accaacgcac tcttctacgt
     2521 cagctccacc atcaacccca tcctgtacaa cctcgtctct gccaacttcc gccacatctt
     2581 cctggccaca ctggcctgcc tctgcccggt gtggcggcgc aggaggaaga ggccagcctt
     2641 ctcgaggaag gccgacagcg tgtccagcaa ccacaccctc tccagcaatg ccacccgcga
     2701 gacgctgtac taggaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     2761 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2821 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2881 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2941 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3001 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3061 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3121 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3181 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3241 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3301 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3361 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3421 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3481 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3541 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3601 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3661 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3721 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3781 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3841 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3901 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3961 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4021 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4081 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4141 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4381 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4441 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4501 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4561 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4621 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4681 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4741 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4801 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4921 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5161 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5461 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5641 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5761 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6601 tgctcacatg t
//
