LOCUS       sequence-266808-        6239 bp    DNA     circular SYN 15-DEC-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 Camk2a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6239)
  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 6239)
  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: "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-pCamk2a-AcGFP
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..451
                     /label=TRE3G BI promoter
                     /note="3rd-generation Tet-responsive bidirectional promoter
                     that can be activated by binding of Tet-On(R) 3G"
     protein_bind    81..99
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    117..135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    225..243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    261..279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    297..315
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    1031..1112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             1547..2266
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     primer_bind     complement(2281..2298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2287..2511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2596..2736
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2596..2725
                     /label=AAV2 ITR
     rep_origin      2811..3266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3108..3129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3283..3302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3402..3424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3462..3480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3653..3721,3722..4513)
                     /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(3871..3890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4684..5272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5173..5192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5334..5463
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    5599..5680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      6218..6227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccac
       61 ctcgacatac tcgagtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc
      121 tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag
      181 gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag
      241 agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc
      301 tatcagtgat agagaacgta taagctttag gcgtgtacgg tgggcgccta taaaagcaga
      361 gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatacc
      421 aactttccgt accacttcct accctcgtaa agtcgacacc ggggcccaga tctatgaagt
      481 tcactgaaat cttccccgtg gaggacgcga actaccctta cagcgccttc atcgcgtcgg
      541 tccggaaaga cgtgatcaaa cactgcaccg accataaagg gatcttccag cccgtgctgc
      601 caccggagaa gaaggtcccg gagctatggt tgtacacaga gctcaaaact aggaccagct
      661 ccatcacgct cgccatacgc atggacaacc tgtacctcgt gggcttcagg accccgggcg
      721 gggtgtggtg ggagttcggc aaggacggcg acacccacct cctcggcgac aaccccaggt
      781 ggctcggctt cggcggcagg taccaggacc tcatcggcaa caagggtctg gagaccgtca
      841 ccatgggccg cgccgaaatg accagggccg tcaacgacct ggcgaagaag aagaagtgag
      901 cggccgccgg cgatatctcc agaggatcat aatcagccat accacatttg tagaggtttt
      961 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1021 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1081 aaatttcaca aataaagcat ttttttcact gccccgagct tcctcgctca ctgactcgct
     1141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggatg tggactaagt
     1201 ttgttcacat ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact
     1261 tgctcctggg cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc
     1321 aggttttaaa gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca
     1381 gggcgtttcg gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt
     1441 gagcagccac agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt
     1501 tgcactcagg agcacgggca ggcgagtggc ccctagttct gggagtatgg tgagcaaggg
     1561 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     1621 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1681 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     1741 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     1801 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     1861 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     1921 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     1981 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     2041 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     2101 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     2161 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     2221 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtgaaacc cgctgatcag
     2281 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2341 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2401 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2461 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2521 cggaaagaac cagatcctct cttaaggtag catcgagatt taaattaggg ataacagggt
     2581 aatggcgcgg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2641 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2701 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2761 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3061 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3121 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3181 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3241 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3301 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3361 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3421 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3481 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3661 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3721 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3781 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3841 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3901 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3961 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4021 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4081 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4141 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4201 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4261 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4321 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4381 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4441 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4501 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4681 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4741 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4801 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4861 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4921 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4981 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5041 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5101 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5161 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5221 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5281 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
     5341 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     5401 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     5461 cctgcggcaa ttcagtcgat aactataacg gtcctaaggt agcgatttaa atacgcgctc
     5521 tcttaaggta gccccgggac gcgtcaattg catgaagaat ctgcttaggg ttaggcgttt
     5581 tgctgccacc tgacgtcggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5641 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5701 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5761 caaatgtggt atggctgatt atgatcctct agactgcaga ctagttcatt gaaagtacaa
     5821 gttctctgaa ccagcttttt tgtagagaga tgttatgctt tcctgggcac aagacattgg
     5881 cagcttgtaa agttcgtcca ttccgtgtgt tatcccagcc gccgcagtag tttgattctt
     5941 aacgagcgca acgatcctcg ctgcttcgtt cttatccttg atgccaagct tctgcatgtc
     6001 ggggatcaca gcggtggggt ggtcagccca ctcgaaggcc gccttggaga tcctgtccca
     6061 cttctgcacc tgcttcccct gcgtcacggt caaggtcacc ccgtgctggc tgttgaaccc
     6121 cgcgtccacc gtgcgggaca cggtgttgaa ccgcagcccc tcgcacacca tgaccaccag
     6181 cttcaccagc ttgctcttcg tgtcggcctg tgggtcagcc gccatggtga attctccag
//
