LOCUS       sequence-266129-        3961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the gePSI-alpha-chain and ZsGreen1 under control of the 
            inducible bi-directional TRE3G promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3961)
  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 3961)
  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-alpha-chain
FEATURES             Location/Qualifiers
     source          1..3961
                     /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    137..155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    173..191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    209..227
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    245..263
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    281..299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    317..335
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    353..371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             466..1161
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    1281..1362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1395..1499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1500..1568,1569..2360)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1718..1737)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2531..3119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3020..3039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3273..3290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3301..3382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tttacgaggg taggaagtgg tacggaaagt tggtataaga caaaagtgtt gtggaattgc
       61 tccaggcgat ctgacggttc actaaacgag ctctgctttt ataggcgccc accgtacacg
      121 cctaaagctt atacgttctc tatcactgat agggagtaaa ctggatatac gttctctatc
      181 actgataggg agtaaactgt agatacgttc tctatcactg atagggagta aactggtcat
      241 acgttctcta tcactgatag ggagtaaact ccttatacgt tctctatcac tgatagggag
      301 taaagtctgc atacgttctc tatcactgat agggagtaaa ctcttcatac gttctctatc
      361 actgataggg agtaaactcg agtatgtcga ggtggcgtgt acggtgggag gcctatataa
      421 gcagagctcg tttagtgaac cgtcagatcg cctggagaat tcaccatggc ccagtccaag
      481 cacggcctga ccaaggagat gaccatgaag taccgcatgg agggctgcgt ggacggccac
      541 aagttcgtga tcaccggcga gggcatcggc taccccttca agggcaagca ggccatcaac
      601 ctgtgcgtgg tggagggcgg ccccttgccc ttcgccgagg acatcttgtc cgccgccttc
      661 atgtacggca accgcgtgtt caccgagtac ccccaggaca tcgtcgacta cttcaagaac
      721 tcctgccccg ccggctacac ctgggaccgc tccttcctgt tcgaggacgg cgccgtgtgc
      781 atctgcaacg ccgacatcac cgtgagcgtg gaggagaact gcatgtacca cgagtccaag
      841 ttctacggcg tgaacttccc cgccgacggc cccgtgatga agaagatgac cgacaactgg
      901 gagccctcct gcgagaagat catccccgtg cccaagcagg gcatcttgaa gggcgacgtg
      961 agcatgtacc tgctgctgaa ggacggtggc cgcttgcgct gccagttcga caccgtgtac
     1021 aaggccaagt ccgtgccccg caagatgccc gactggcact tcatccagca caagctgacc
     1081 cgcgaggacc gcagcgacgc caagaaccag aagtggcacc tgaccgagca cgccatcgcc
     1141 tccggctccg ccttgccctg actgcagtct agaggatcat aatcagccat accacatttg
     1201 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1261 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1321 atagcatcac aaatttcaca aataaagcat ttttttcact gccgacgtca ggtggcactt
     1381 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1441 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1501 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1561 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1621 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1681 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1741 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1801 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1861 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1921 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1981 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2041 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2101 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2161 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2221 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2281 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2341 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2401 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2461 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2521 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2581 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2641 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2701 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2761 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2821 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2881 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2941 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3001 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3061 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3121 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3181 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3241 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagctcggg
     3301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3421 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     3481 ttatgatcct ctggagatat cgccggcggc cgctcacttc ttcttcttcg ccaggtcgtt
     3541 gacggccctg gtcatttcgg cgcggcccat ggtgacggtc tccagaccct tgttgccgat
     3601 gaggtcctgg tacctgccgc cgaagccgag ccacctgggg ttgtcgccga ggaggtgggt
     3661 gtcgccgtcc ttgccgaact cccaccacac cccgcccggg gtcctgaagc ccacgaggta
     3721 caggttgtcc atgcgtatgg cgagcgtgat ggagctggtc ctagttttga gctctgtgta
     3781 caaccatagc tccgggacct tcttctccgg tggcagcacg ggctggaaga tccctttatg
     3841 gtcggtgcag tgtttgatca cgtctttccg gaccgacgcg atgaaggcgc tgtaagggta
     3901 gttcgcgtcc tccacgggga agatttcagt gaacttcata gatctgggcc ccggtgtcga
     3961 c
//
