LOCUS       sequence-337500-        5648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SST interneuron-restricted gene regulatory element GRE22, to drive 
            GFP expression in SST+ interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Hrvatin S, Tzeng CP, Nagy MA, Stroud H, Koutsioumpa C, Wilcox OF, 
            Assad EG, Green J, Harvey CD, Griffith EC, Greenberg ME
  TITLE     A scalable platform for the development of cell-type-specific viral 
            drivers.
  JOURNAL   Elife. 2019 Sep 23;8. pii: 48089. doi: 10.7554/eLife.48089.
  PUBMED    31545165
REFERENCE   2  (bases 1 to 5648)
  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: "Elife."; 
            date: "2019-09-23"; volume: "8. pii: 48089. doi"; pages: " 
            10.7554/eLife.48089"
COMMENT     Sequence Label: pAAV-Sst22-GFP
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      449..1037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     938..957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1099..1228
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          2132..2264
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2247..2266
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2307..2316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2313..2315,2316..2318,2319..3032)
                     /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(2313..2315,2316..2318,2319..3029)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2358..2379)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2619..2638)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2966..2987
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3077..3665
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3130..3150)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3548..3559)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3849..3930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3886..3905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3940..3959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4009..4149
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4009..4138
                     /label=AAV2 ITR
     rep_origin      4224..4679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4311..4330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4521..4542
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4696..4715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4815..4837
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4875..4893)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4961..5065
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5066..5134,5135..5648,1..278)
                     /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(5284..5303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
       61 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
      121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      241 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
      301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
      361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
      421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
      601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1081 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     1141 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     1201 gtggccaact ccatcactag gggttcctgc ggccgcacgc gtttaatagc caggactaca
     1261 cagagaaacc ctgtctcaaa aaaacaaaat caaaacaaaa caaacaaaca aaaaagctaa
     1321 tgactccatc atgactgtaa caaacacatc agtgcggcag tgagagcccg tctgtcagca
     1381 tcagcaacag cattagtcag actgtatttg tgagcatatt tgcttaggtc tcttctaaat
     1441 acccttcact tttctctcag agaaacccag ttcatcgtat tctgaaaagg agcggccgta
     1501 aaggactgat cctgtctgaa gcactttggt ataaaagttg cttagcagtg gggcagaaaa
     1561 gaaaaaaagc aattaagttt atatttagtg atctatctat acacatctgg agcacatttg
     1621 ggaaagaatt caaaagggcc aattcattgc atgcctcctg ctacagaacg agtgtgggag
     1681 tcaagctgcg atttccacag catcagacat ttattgttga cttcaaaaag ttctcccact
     1741 tatgtgtaat tactatccta gcaaatggct ctgaaatttc agcttcttaa gcataaggca
     1801 gagtggtcct ttaaaagtaa aataaaacgt aggccctatg agataaaatt aagataaatt
     1861 aagaatcagt tacttccaag acgaagcact tatggtgcat gccttcttat ataaagcaga
     1921 tccttaccat gtatgtgtgc tgtttgcttg ccaagaccaa gatgtctgtc gactgatatc
     1981 gaattcctgc agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     2041 tgcttctagc ctgcaggtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag
     2101 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
     2161 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     2221 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
     2281 caattacagc tcttaagaaa ctagtagcca ccatggtgag caagggcgag gagctgttca
     2341 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2401 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2461 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     2521 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2581 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2641 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2701 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2761 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2821 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2881 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2941 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3001 tcactctcgg catggacgag ctgtacaagt aaaggcgcgc cacccctgca gggaattcga
     3061 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3121 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3181 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3241 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3301 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3361 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3421 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3481 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     3541 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3601 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3661 cccgcatcga taccgagcgc gcgatcgcaa tcggagaatc gagagatctg tgatagcggc
     3721 catcaagctg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3781 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3841 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3901 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3961 atgtatcagc ttatcgatac cgcatgcacg tgcggaccga gcggccgcag gaacccctag
     4021 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4081 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4141 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4201 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4261 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4321 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4381 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4441 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4501 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4561 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4621 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4681 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4741 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4801 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4861 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4921 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4981 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5041 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5101 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5161 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5221 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5281 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5341 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5401 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5461 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5521 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5581 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5641 aactggcg
//
