LOCUS       sequence-337494-        5673 bp    DNA     circular SYN 14-APR-2025
DEFINITION  SST interneuron-restricted gene regulatory element GRE12, to drive 
            GFP expression in SST+ interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5673)
  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 5673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Sst12-GFP
FEATURES             Location/Qualifiers
     source          1..5673
                     /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          2158..2290
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2273..2292
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2333..2342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2339..2341,2342..2344,2345..3058)
                     /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(2339..2341,2342..2344,2345..3055)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2384..2405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2645..2664)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2992..3013
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3103..3691
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3156..3176)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3574..3585)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3874..3955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3911..3930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3965..3984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4034..4174
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4034..4163
                     /label=AAV2 ITR
     rep_origin      4249..4704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4336..4355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4546..4567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4721..4740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4840..4862
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4900..4918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4986..5090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5091..5159,5160..5673,1..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(5309..5328)
                     /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 gtttaatctt tagaggggga
     1261 aactgccttt tgagttgttt atatataaag ttatttaaat aatgaagatc atttttttct
     1321 gcctataatg tttttcttga gatgatgctt tcttgaaaaa aatattttca aaggctgaaa
     1381 acaaatacat aagaactcag taaactcggg aagtgtttag cttcataatc agactgtgca
     1441 gaagatagga agcagcagcc ggatccacag cctctgattg tcccaaatca caggagtcat
     1501 caactgagta ctccaaaaag gaaaacaagc cattttcagc taaaagatat gagcataatg
     1561 tgtaccataa tctcacagtg gctgttttag aaccaagagt gtttgtgact taatttgaat
     1621 ttctcaatgc aacatttctc aaaaattcct taaacgtcat gtcatagatg atttattatg
     1681 tacaaaacat aactgttgag aaactccatt tccttgcctt ctgggaggaa ccttaggaaa
     1741 catcagcagc aggtgcaaag tattccatag agagagggct ggcataaaga acatatttat
     1801 tcatcagttc caaatttccc tgcttctgag ggcttaaaaa gagggatttc ttgagctgag
     1861 gaaattaaaa acaaaacaaa caactatgct gaaagaggac tagaaatgtt ctgggatatt
     1921 gtgaaatcta gacttgaaat tccttctcat ttccttatgc acagatttta acacccttgg
     1981 tttcttcgga gtagtcgact gatatcgaat tcctgcagcc cgggctgggc ataaaagtca
     2041 gggcagagcc atctattgct tacatttgct tctagcctgc aggtcgagga gcgcagcctt
     2101 ccagaagcag agcgcggcgc cttaagctgc agaagttggt cgtgaggcac tgggcaggta
     2161 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     2221 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     2281 ctctccacag gtgtccactc ccagttcaat tacagctctt aagaaactag tagccaccat
     2341 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2401 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2461 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2521 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2581 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2641 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2701 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2761 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2821 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2881 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2941 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3001 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     3061 gcgcgccacc cctgcaggga attcgatatc aagcttatcg ataatcaacc tctggattac
     3121 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3181 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3241 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3301 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3361 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3421 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3481 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     3541 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3601 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3661 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgcgcga tcgcatatgg
     3721 ctgtcgagag atctgtgata gcggccatca agctggccgc gactctagat cataatcagc
     3781 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3841 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3961 agttgtggtt tgtccaaact catcaatgta tcagcttatc gataccgcat gcacgtgcgg
     4021 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4081 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4141 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4201 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4261 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4321 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4381 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4441 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4501 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4561 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4621 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4681 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4741 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4801 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4861 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4921 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5101 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5161 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5221 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5281 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5341 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5401 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5461 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5521 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5581 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5641 tggcaacaac gttgcgcaaa ctattaactg gcg
//
