LOCUS       sequence-436531-        6486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split intein version of Trojan-Gal4. Create Gal4 driver lines from 
            MiMIC lines, in which MiMIC cassette was inserted into Phase 1 
            coding intron. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  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 6486)
  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(1)-Gal4
FEATURES             Location/Qualifiers
     source          1..6486
                     /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             583..636
                     /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             637..3279
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4"
                     /product="GAL4 transcriptional activator"
                     /label=GAL4
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
                     MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
                     RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
                     LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
                     KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
                     IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
                     KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
                     RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
                     NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
                     QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
                     VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
                     QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             3280..3333
                     /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    3585..3654
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(3700..3720)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3700..3718)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3739..3755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3739..3755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3752..3774)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3763..3779
                     /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(3787..3793,3794..3811,3812..3817))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3832..3853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3970..3987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4141..4729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4221..4240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4900..5691,5692..5760))
                     /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     5523..5542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5761..5865)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5892..6347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5979..5998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6189..6210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6474..6486,1..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 gagtcgatcc
      181 aacatggcga cttgtcccat ccccggcatg tttaaatata ctaattattc ttgaactaat
      241 tttaatcaac cgatttatct ctcttccgca ggtgaatatt gcctgtccta cgacaccgag
      301 atcctgaccg tggagtacgg cttcctgccc atcggcaaga tcgtggagga gcgcatcgag
      361 tgcaccgtgt acaccgtgga caagaacggc ttcgtgtaca cccagcccat cgcccagtgg
      421 cacaaccgcg gcgagcagga ggtgttcgag tactgcctgg aggacggctc catcatccgc
      481 gccaccaagg accacaagtt catgaccacc gacggccaga tgctgcccat cgacgagatc
      541 ttcgagcgcg gcctggacct gaagcaggtg gacggcctgc ccgagggccg cggcagcctg
      601 ctgacctgcg gcgatgtgga ggagaacccc gggcccatga agctactgtc ttctatcgaa
      661 caagcctgcg atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc
      721 gccaagtgtc tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg
      781 ctgactaggg cacatctgac agaagtggaa tcaaggctag aaagactgga acagctattt
      841 ctactgattt ttcctcgaga agaccttgac atgattttga aaatggattc tttacaggat
      901 ataaaagcat tgttaacagg attatttgta caagataatg tgaataaaga tgccgtcaca
      961 gatagattgg cttcagtgga gactgatatg cctctaacat tgagacagca tagaataagt
     1021 gcgacatcat catcggaaga gagtagtaac aaaggtcaaa gacagttgac tgtatcgatt
     1081 gactcggcag ctcatcatga taactccaca attccgttgg attttatgcc cagggatgct
     1141 cttcatggat ttgattggtc tgaagaggat gacatgtcgg atggcttgcc cttcctgaaa
     1201 acggacccca acaataatgg gttctttggc gacggttctc tcttatgtat tcttcgatct
     1261 attggcttta aaccggaaaa ttacacgaac tctaacgtta acaggctccc gaccatgatt
     1321 acggatagat acacgttggc ttctagatcc acaacatccc gtttacttca aagttatctc
     1381 aataattttc acccctactg ccctatcgtg cactcaccga cgctaatgat gttgtataat
     1441 aaccagattg aaatcgcgtc gaaggatcaa tggcaaatcc tttttaactg catattagcc
     1501 attggagcct ggtgtataga gggggaatct actgatatag atgtttttta ctatcaaaat
     1561 gctaaatctc atttgacgag caaggtcttc gagtcaggtt ccataatttt ggtgacagcc
     1621 ctacatcttc tgtcgcgata tacacagtgg aggcagaaaa caaatactag ctataatttt
     1681 cacagctttt ccataagaat ggccatatca ttgggcttga atagggacct cccctcgtcc
     1741 ttcagtgata gcagcattct ggaacaaaga cgccgaattt ggtggtctgt ctactcttgg
     1801 gagatccaat tgtccctgct ttatggtcga tccatccagc tttctcagaa tacaatctcc
     1861 ttcccttctt ctgtcgacga tgtgcagcgt accacaacag gtcccaccat atatcatggc
     1921 atcattgaaa cagcaaggct cttacaagtt ttcacaaaaa tctatgaact agacaaaaca
     1981 gtaactgcag aaaaaagtcc tatatgtgca aaaaaatgct tgatgatttg taatgagatt
     2041 gaggaggttt cgagacaggc accaaagttt ttacaaatgg atatttccac caccgctcta
     2101 accaatttgt tgaaggaaca cccttggcta tcctttacaa gattcgaact gaagtggaaa
     2161 cagttgtctc ttatcattta tgtattaaga gattttttca ctaattttac ccagaaaaag
     2221 tcacaactag aacaggatca aaatgatcat caaagttatg aagttaaacg atgctccatc
     2281 atgttaagcg atgcagcaca aagaactgtt atgtctgtaa gtagctatat ggacaatcat
     2341 aatgtcaccc catattttgc ctggaattgt tcttattact tgttcaatgc agtcctagta
     2401 cccataaaga ctctactctc aaactcaaaa tcgaatgctg agaataacga gaccgcacaa
     2461 ttattacaac aaattaacac tgttctgatg ctattaaaaa aactggccac ttttaaaatc
     2521 cagacttgtg aaaaatacat tcaagtactg gaagaggtat gtgcgccgtt tctgttatca
     2581 cagtgtgcaa tcccattacc gcatatcagt tataacaata gtaatggtag cgccattaaa
     2641 aatatcgtcg gttctgcaac tatcgcccaa taccctactc ttccggagga aaatgtcaac
     2701 aatatcagtg ttaaatatgt ttctcctggc tcagtagggc cttcacctgt gccattgaaa
     2761 tcaggagcaa gtttcagtga tctagtcaag ctgttatcta accgtccacc ctctcgtaac
     2821 tctccagtga caataccaag aagcacacct tcgcatcgct cagtcacgcc ttttctaggg
     2881 caacagcaac agctgcaatc attagtgcca ctgaccccgt ctgctttgtt tggtggcgcc
     2941 aattttaatc aaagtgggaa tatcgctgat agctcattgt ccttcacttt cactaacagt
     3001 agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc acaaccaatt
     3061 gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag taaaattgat
     3121 gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc gtataacgcg
     3181 tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata taactatcta
     3241 ttcgatgatg aagatacccc accaaaccca aaaaaagagg agggccgcgg cagcctgctg
     3301 acctgcggcg atgtggagga gaaccccggg cccgtgaaga tcatctcccg caagtccctg
     3361 ggcacccaga acgtgtacga catcggcgtg gagaaggacc acaacttcct gctgaagaac
     3421 ggcctggtgg cctccaactg cttcaacggt aagttattga acaatggcat caaatgcctt
     3481 catcatcact accctttagc ccttaagacc ccacaatgac cttacccact cagagaaaaa
     3541 agtaaatatg aaagcccatt tgaacttctc atcatgatgg accagatggg tgaggtggag
     3601 tacgcgcccg gggagcccaa gggcacgccc tggcacccgc accgcggctt cgagaccgtg
     3661 acctacatcg gcgcgccggt accactagtg gggaggttcc ctttagtgag ggttaattgc
     3721 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3781 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3841 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3901 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4561 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5881 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5941 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6241 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6301 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt
     6361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6481 acgacg
//
