LOCUS       sequence-266117-        3847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the gePSI-beta-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 3847)
  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 3847)
  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-beta-chain
FEATURES             Location/Qualifiers
     source          1..3847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    43..61
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    79..97
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    115..133
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    151..169
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             264..959
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    1079..1160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1193..1297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1298..1366,1367..2158)
                     /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(1516..1535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2329..2917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2818..2837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3071..3088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3099..3180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        join(3646..3847,1..249)
                     /label=TRE3G BI promoter
                     /note="3rd-generation Tet-responsive bidirectional promoter
                     that can be activated by binding of Tet-On(R) 3G"
     protein_bind    3782..3800
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3818..3836
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 atacgttctc tatcactgat agggagtaaa ctggtcatac gttctctatc actgataggg
       61 agtaaactcc ttatacgttc tctatcactg atagggagta aagtctgcat acgttctcta
      121 tcactgatag ggagtaaact cttcatacgt tctctatcac tgatagggag taaactcgag
      181 tatgtcgagg tggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg
      241 tcagatcgcc tggagaattc accatggccc agtccaagca cggcctgacc aaggagatga
      301 ccatgaagta ccgcatggag ggctgcgtgg acggccacaa gttcgtgatc accggcgagg
      361 gcatcggcta ccccttcaag ggcaagcagg ccatcaacct gtgcgtggtg gagggcggcc
      421 ccttgccctt cgccgaggac atcttgtccg ccgccttcat gtacggcaac cgcgtgttca
      481 ccgagtaccc ccaggacatc gtcgactact tcaagaactc ctgccccgcc ggctacacct
      541 gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat ctgcaacgcc gacatcaccg
      601 tgagcgtgga ggagaactgc atgtaccacg agtccaagtt ctacggcgtg aacttccccg
      661 ccgacggccc cgtgatgaag aagatgaccg acaactggga gccctcctgc gagaagatca
      721 tccccgtgcc caagcagggc atcttgaagg gcgacgtgag catgtacctg ctgctgaagg
      781 acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa ggccaagtcc gtgccccgca
      841 agatgcccga ctggcacttc atccagcaca agctgacccg cgaggaccgc agcgacgcca
      901 agaaccagaa gtggcacctg accgagcacg ccatcgcctc cggctccgcc ttgccctgac
      961 tgcagtctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1021 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1141 taaagcattt ttttcactgc cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1201 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1261 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1321 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1381 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1441 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1501 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1561 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1621 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1681 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1741 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1801 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1861 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1921 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1981 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2041 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2101 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2161 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2221 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2281 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2341 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2401 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2461 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2521 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2581 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2641 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2701 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2761 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2821 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2881 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2941 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3001 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3061 gaccgagcgc agcgagtcag tgagcgagga agctcggggc agtgaaaaaa atgctttatt
     3121 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3181 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3241 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct ggagatatcg
     3301 ccggcggccg ctcaagccgc cgcagtagtt tgattcttaa cgagcgcaac gatcctcgct
     3361 gcttcgttct tatccttgat gccaagcttc tgcatgtcgg ggatcacagc ggtggggtgg
     3421 tcagcccact cgaaggccgc cttggagatc ctgtcccact tctgcacctg cttcccctgc
     3481 gtcacggtca aggtcacccc gtgctggctg ttgaaccccg cgtccaccgt gcgggacacg
     3541 gtgttgaacc gcagcccctc gcacaccatg accaccagct tcaccagctt gctcttcgtg
     3601 tcggcctgtg ggtcagccgc catagatctg ggccccggtg tcgactttac gagggtagga
     3661 agtggtacgg aaagttggta taagacaaaa gtgttgtgga attgctccag gcgatctgac
     3721 ggttcactaa acgagctctg cttttatagg cgcccaccgt acacgcctaa agcttatacg
     3781 ttctctatca ctgataggga gtaaactgga tatacgttct ctatcactga tagggagtaa
     3841 actgtag
//
