LOCUS       sequence-337508-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SST interneuron-restricted gene regulatory element GRE44, to drive 
            GFP expression in SST+ interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  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 5647)
  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-Sst44-GFP
FEATURES             Location/Qualifiers
     source          1..5647
                     /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          2131..2263
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2246..2265
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2306..2315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2312..2314,2315..2317,2318..3031)
                     /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(2312..2314,2315..2317,2318..3028)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2357..2378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2618..2637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2965..2986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3076..3664
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3129..3149)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3547..3558)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3848..3929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3885..3904)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3939..3958
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4008..4148
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4008..4137
                     /label=AAV2 ITR
     rep_origin      4223..4678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4310..4329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4520..4541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4695..4714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4814..4836
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4874..4892)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4960..5064
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5065..5133,5134..5647,1..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(5283..5302)
                     /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 gtttaatgtt cagtacccag
     1261 acactcccat acccttattt agaagaaata aatatcatca agtcataata tccttgactg
     1321 attaagaaag ccactttgta agtgtttatt aaactgtcaa gaaacttaca gaatttacta
     1381 catgatcgtt agaataactt tgagtcagga catatttgat atgacttaat catactccct
     1441 ccaaaaggaa ataaggcttt gtgaaggtaa attatttctt cctgggttgg atatgtgttt
     1501 atggagtgat cattcagctg ttcccaacct tcattctgaa aaggcctcag aacacttcat
     1561 gatgaatcaa gctgtatcct gaatagagta aaatgaacca cttcgtagga actatggtgt
     1621 caccacatca gcaattctta ttgaaaagtg tgcatttctt attcacatat ttcaaagatg
     1681 gtattccaga ggagtgattt tctcaatgta tttttcatct acaagccttc attttaagcc
     1741 taccaccgtg tgtgttttca agacagcaat tatcgtttta aaatgtgcag gtctagcttg
     1801 agcttctcag caagtttcta tgccaaagaa aacaccaatc ctttccattt actgagaatc
     1861 aatgtttaat cctccttttt gttctcatac ttattacaaa tcataaagaa ttctgagtgt
     1921 cagtttgata actagaagct ccatgtacca ttcctgctcc ttattgagtc gactgtatcg
     1981 aattcctgca gcccgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     2041 gcttctagcc tgcaggtcga ggagcgcagc cttccagaag cagagcgcgg cgccttaagc
     2101 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
     2161 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     2221 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc
     2281 aattacagct cttaagaaac tagtagccac catggtgagc aagggcgagg agctgttcac
     2341 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2401 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2461 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2521 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2581 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2641 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2701 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2761 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2821 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2881 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2941 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3001 cactctcggc atggacgagc tgtacaagta aaggcgcgcc acccctgcag ggaattcgat
     3061 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3121 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3181 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3241 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3301 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3361 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3421 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3481 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3541 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3601 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3661 ccgcatcgat accgagcgcg cgatcgctaa tggccaatcg agagatctgt gatagcggcc
     3721 atcaagctgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3781 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3841 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3961 tgtatcagct tatcgatacc gcatgcacgt gcggaccgag cggccgcagg aacccctagt
     4021 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4081 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4141 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4201 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4261 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4321 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4381 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4441 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4501 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4561 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4621 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4681 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4741 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4801 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4861 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4921 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4981 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5041 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5101 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5161 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5221 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5281 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5341 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5401 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5461 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5521 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5581 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5641 actggcg
//
