LOCUS       sequence-265762-        7559 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses both the gePSI-alpha-chain and the gePSI-beta-chain under 
            control of the inducible bi-directional TRE3G promoter. Expresses 
            constitutively AcGFP under control of a GFAP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  AUTHORS   Heumuller M, Glock C, Rangaraju V, Biever A, Schuman EM
  TITLE     A genetically encodable cell-type-specific protein synthesis 
            inhibitor.
  JOURNAL   Nat Methods. 2019 Jul 15. pii: 10.1038/s41592-019-0468-x. doi: 
            10.1038/s41592-019-0468-x.
  PUBMED    31308551
REFERENCE   2  (bases 1 to 7559)
  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: "Nat 
            Methods. 2019 Jul 15. pii: 10.1038/s41592-019-0468-x. doi: 
            10.1038/s41592-019-0468-x."
COMMENT     Sequence Label: pTRE3G-Bi-gePSI-pGFAP-AcGFP
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   36..165
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    301..382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      920..929
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        942..1392
                     /label=TRE3G BI promoter
                     /note="3rd-generation Tet-responsive bidirectional promoter
                     that can be activated by binding of Tet-On(R) 3G"
     protein_bind    1022..1040
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1058..1076
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1094..1112
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1130..1148
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1166..1184
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1202..1220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1238..1256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    1972..2053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             3808..4527
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     primer_bind     complement(4542..4559)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4548..4772
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4857..4997
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4857..4986
                     /label=AAV2 ITR
     rep_origin      5072..5527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5159..5178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5369..5390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5544..5563)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5663..5685
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5723..5741)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5809..5913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5914..5982,5983..6774)
                     /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(6132..6151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6945..7533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7434..7453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
       61 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
      121 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc aattcagtcg
      181 ataactataa cggtcctaag gtagcgattt aaatacgcgc tctcttaagg tagccccggg
      241 acgcgtcaat tgcatgaaga atctgcttag ggttaggcgt tttgctgcca cctgacgtcg
      301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      421 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
      481 ttatgatcct ctagactgca gactagttca ttgaaagtac aagttctctg aaccagcttt
      541 tttgtagaga gatgttatgc tttcctgggc acaagacatt ggcagcttgt aaagttcgtc
      601 cattccgtgt gttatcccag ccgccgcagt agtttgattc ttaacgagcg caacgatcct
      661 cgctgcttcg ttcttatcct tgatgccaag cttctgcatg tcggggatca cagcggtggg
      721 gtggtcagcc cactcgaagg ccgccttgga gatcctgtcc cacttctgca cctgcttccc
      781 ctgcgtcacg gtcaaggtca ccccgtgctg gctgttgaac cccgcgtcca ccgtgcggga
      841 cacggtgttg aaccgcagcc cctcgcacac catgaccacc agcttcacca gcttgctctt
      901 cgtgtcggcc tgtgggtcag ccgccatggt gaattctcca ggcgatctga cggttcacta
      961 aacgagctct gcttatatag gcctcccacc gtacacgcca cctcgacata ctcgagttta
     1021 ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt
     1081 atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt
     1141 gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta
     1201 ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt
     1261 ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc
     1321 agatcgcctg gagcaattcc acaacacttt tgtcttatac caactttccg taccacttcc
     1381 taccctcgta aagtcgacac cggggcccag atctatgaag ttcactgaaa tcttccccgt
     1441 ggaggacgcg aactaccctt acagcgcctt catcgcgtcg gtccggaaag acgtgatcaa
     1501 acactgcacc gaccataaag ggatcttcca gcccgtgctg ccaccggaga agaaggtccc
     1561 ggagctatgg ttgtacacag agctcaaaac taggaccagc tccatcacgc tcgccatacg
     1621 catggacaac ctgtacctcg tgggcttcag gaccccgggc ggggtgtggt gggagttcgg
     1681 caaggacggc gacacccacc tcctcggcga caaccccagg tggctcggct tcggcggcag
     1741 gtaccaggac ctcatcggca acaagggtct ggagaccgtc accatgggcc gcgccgaaat
     1801 gaccagggcc gtcaacgacc tggcgaagaa gaagaagtga gcggccgccg gcgatatctc
     1861 cagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1921 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2041 tttttttcac tgccccgagc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2101 cggcgagcgg tatcagctca ctcaaagggt ctgtaagctg aagacctggc agtgctgagc
     2161 tggtcagccc ccaggacctc cttttgtgcc cacgagtgaa tcaccttggc atagacataa
     2221 tggtcagggg tgggcacgca gcctgcttcc cgctgtgctc caggcctcct tcgatgcttt
     2281 ccgagaagtc tattgagctg ggagcttgta ctgcacccgg ggctgacatc ctggcatcct
     2341 gggataaaag cagcccacgg ggctgccctt gccatatgcc tcactggcgg cagagaacaa
     2401 ggctctattc agcgagtacc ctggagtaga caccagaagc ccaagcatgg gcagaggaag
     2461 gcaggggttg gggggagcag agctgtctgt gttccagaag cccaaggaca cagatggcta
     2521 aggcgcctgg gagagggacc tgagtggaag agatagatgg gcctgaagtc tcaagcagca
     2581 acagcctcct ccccgccatt ggtgagggtg gggtttggtt tcccggacct acatatccct
     2641 cagaggcctg gtgtgtagga atttaaaggg ggtaaatctc ctgagagaat gaggggtacc
     2701 caggaagacg gggtgttaca gaaagaaaga ctccagcatg cacagccaac tcattcaaaa
     2761 ctactctgtc aggggctgcc aggggccagg ctcggggtgg ggggtggggg gcaacgagaa
     2821 gctggatcag ggagaaatgg cccactaggc tggataagag gccacagagg ggctcaggaa
     2881 tgaagcctgc tgtcttaccc tattaggatc tgcgtgcata ccttctgccg tgcactctaa
     2941 acacacagcc agaggctcaa gttgaccctg gagtcacaga gagggctcca accttagccc
     3001 tccactcctg aactccagga atgagaagat agagttggag agattcaggg gagaggactc
     3061 tgttgagaat gggggtcaca ggaaactgta atataggttg atcccggagg aagggaatag
     3121 gttcttcaag ttcctagcat ctcacaggcc cccagagaag gacagagttg gggtggtcct
     3181 ggcttacagg ctctaagaac tggaagctga ttaccccacc gagctgtgca ctctctgtct
     3241 ctgtctctgt gtgtgcgctc gtgcacactt atcacacaaa tgttcatgtg tgtgcacata
     3301 catgtgttga gaccagaggt caacctcagg cactgttgcc ttggttttct gagagagcat
     3361 ttctctctgg atctggaact cgccaattag tgagagccag gaagtctgct gattttcact
     3421 gcccagcact ggagtttaca agtatgcact gtcaacccag gccttttgta ttcattctgc
     3481 agctagaact tgggtgggtc ttcatgcttg acaggcaagc aatttatgga ctaagctgtt
     3541 ccctcggccc tctcttgacc catttaccag aaagggggtt ccttgatcaa tggcgaagcc
     3601 aggctggtgt tcccaagaaa gccttgactc tgggtacagt gacctcagtg gggtgagagg
     3661 agttctcccc ctagctgggc tggggcccag ctccaccccc tcaggctatt caatgggggt
     3721 gcttccagga agtcaggggc agatttagtc caacccgttc ctccataaag gccctgacat
     3781 cccaggagcc agcagaggca gggcaccatg gtgagcaagg gcgccgagct gttcaccggc
     3841 atcgtgccca tcctgatcga gctgaatggc gatgtgaatg gccacaagtt cagcgtgagc
     3901 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     3961 ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc tgagctacgg cgtgcagtgc
     4021 ttctcacgct accccgatca catgaagcag cacgacttct tcaagagcgc catgcctgag
     4081 ggctacatcc aggagcgcac catcttcttc gaggatgacg gcaactacaa gtcgcgcgcc
     4141 gaggtgaagt tcgagggcga taccctggtg aatcgcatcg agctgaccgg caccgatttc
     4201 aaggaggatg gcaacatcct gggcaataag atggagtaca actacaacgc ccacaatgtg
     4261 tacatcatga ccgacaaggc caagaatggc atcaaggtga acttcaagat ccgccacaac
     4321 atcgaggatg gcagcgtgca gctggccgac cactaccagc agaatacccc catcggcgat
     4381 ggccctgtgc tgctgcccga taaccactac ctgtccaccc agagcgccct gtccaaggac
     4441 cccaacgaga agcgcgatca catgatctac ttcggcttcg tgaccgccgc cgccatcacc
     4501 cacggcatgg atgagctgta caagtgaaac ccgctgatca gcctcgactg tgccttctag
     4561 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4621 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4681 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4741 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagatcctc
     4801 tcttaaggta gcatcgagat ttaaattagg gataacaggg taatggcgcg ggccgcagga
     4861 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4921 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4981 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5041 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     5101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     5161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5281 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5401 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     5461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5521 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5581 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5641 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5701 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5761 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5881 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5941 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6001 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6061 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6121 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6181 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6241 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6301 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6361 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6421 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6481 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6541 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6601 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6661 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6721 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6781 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6841 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6901 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6961 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7021 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7081 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7141 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7201 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7261 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7321 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7381 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7441 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7501 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacg
//
