LOCUS       sequence-436806-        4476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split intein version of Trojan-Gal4DBD. Create Gal4DBD hemi-driver 
            lines from MiMIC lines, in which MiMIC cassette was inserted into 
            Phase 2 coding intron. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4476)
  AUTHORS   Diao F, Vasudevan D, Heckscher ES, White BH
  TITLE     Hox gene-specific cellular targeting using split intein Trojan 
            exons.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Apr 23;121(17):e2317083121. doi: 
            10.1073/pnas.2317083121. Epub 2024 Apr 11.
  PUBMED    38602904
REFERENCE   2  (bases 1 to 4476)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-04-23"; volume: "121(17):e2317083121. 
            doi"; pages: " 10.1073/pnas.2317083121. Epub 2024 Apr 11"
COMMENT     Sequence Label: psiTE(2)-Gal4DBD
FEATURES             Location/Qualifiers
     source          1..4476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..19
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3..19
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    87..156
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             588..641
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             828..1268
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1269..1322
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    1575..1644
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(1690..1710)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1690..1708)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1729..1745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1729..1745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1742..1764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1753..1769
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1777..1783,1784..1801,1802..1807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1822..1843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1960..1977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2131..2719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2211..2230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2890..3681,3682..3750))
                     /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     3513..3532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3751..3855)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3882..4337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3969..3988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4179..4200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(4464..4476,1..10)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgacc caccggtgca
       61 tgcgcggccg cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc
      121 ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat gtctagaagt
      181 cgatccaaca tggcgacttg tcccatcccc ggcatgttta aatatactaa ttattcttga
      241 actaatttta atcaaccgat ttatctctct tccgcaggga atattgcctg tcctacgaca
      301 ccgagatcct gaccgtggag tacggcttcc tgcccatcgg caagatcgtg gaggagcgca
      361 tcgagtgcac cgtgtacacc gtggacaaga acggcttcgt gtacacccag cccatcgccc
      421 agtggcacaa ccgcggcgag caggaggtgt tcgagtactg cctggaggac ggctccatca
      481 tccgcgccac caaggaccac aagttcatga ccaccgacgg ccagatgctg cccatcgacg
      541 agatcttcga gcgcggcctg gacctgaagc aggtggacgg cctgcccgag ggccgcggca
      601 gcctgctgac ctgcggcgat gtggaggaga accccgggcc catgctggag atccgcgccg
      661 ccttcctgcg ccagcgcaac accgccctgc gcaccgaggt ggccgagctg gagcaggagg
      721 tgcagcgcct ggagaacgag gtgagccagt acgagacccg ctacggcccc ctgggcggcg
      781 gcaaggctag cggaggaggt ggtggaggtg gaggtggagg tactagtatg aagctgctga
      841 gtagtattga acaagcgtgt gacatatgcc gacttaagaa gctgaagtgc agcaaggaaa
      901 agccgaagtg cgccaaatgt cttaagaaca attgggagtg ccggtactcg ccaaagacga
      961 agcgaagccc tttgacacgc gctcatttga ctgaagtgga gagtcgcctg gagcgccttg
     1021 agcagttgtt cctcctcatt tttccccgtg aagacctcga catgatactg aagatggata
     1081 gtctccaaga tattaaggcc ctgctcactg gcctgtttgt tcaggataac gtcaacaagg
     1141 atgccgtgac ggaccgcctt gcctcggtgg agacagatat gccacttacc ttgcggcagc
     1201 atcgcatctc ggctaccagc agtagtgaag aaagttccaa taagggtcaa cggcaactga
     1261 cggtatcgga aggacgcgga tcgttgctga cgtgcggaga cgtggaggag aatccgggcc
     1321 ccgtgaagat catctcccgc aagtccctgg gcacccagaa cgtgtacgac atcggcgtgg
     1381 agaaggacca caacttcctg ctgaagaacg gcctggtggc ctccaactgc ttcaacaggt
     1441 aagttattga acaatggcat caaatgcctt catcatcact accctttagc ccttaagacc
     1501 ccacaatgac cttacccact cagagaaaaa agtaaatatg aaagcccatt tgaacttctc
     1561 atcatgatgg accagatggg tgaggtggag tacgcgcccg gggagcccaa gggcacgccc
     1621 tggcacccgc accgcggctt cgagaccgtg acctacatcg gcgcgccggt accactagtg
     1681 gggaggttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     1741 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1801 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1861 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4201 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4321 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4381 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4441 aagttgggta acgccagggt tttcccagtc acgacg
//
