LOCUS       sequence-436787-        6492 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 0 
            coding intron. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  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 6492)
  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(0)-Gal4
FEATURES             Location/Qualifiers
     source          1..6492
                     /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             587..640
                     /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             641..3283
                     /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             3284..3337
                     /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    3591..3660
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(3706..3726)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3706..3724)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3745..3761)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3745..3761)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3758..3780)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3769..3785
                     /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(3793..3799,3800..3817,3818..3823))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3838..3859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3976..3993)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4147..4735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4227..4246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4906..5697,5698..5766))
                     /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     5529..5548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5767..5871)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5898..6353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5985..6004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6195..6216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6480..6492,1..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 tccgcaggaa tattgcctgt cctacgacac
      301 cgagatcctg accgtggagt acggcttcct gcccatcggc aagatcgtgg aggagcgcat
      361 cgagtgcacc gtgtacaccg tggacaagaa cggcttcgtg tacacccagc ccatcgccca
      421 gtggcacaac cgcggcgagc aggaggtgtt cgagtactgc ctggaggacg gctccatcat
      481 ccgcgccacc aaggaccaca agttcatgac caccgacggc cagatgctgc ccatcgacga
      541 gatcttcgag cgcggcctgg acctgaagca ggtggacggc ctgcccgagg gccgcggcag
      601 cctgctgacc tgcggcgatg tggaggagaa ccccgggccc atgaagctac tgtcttctat
      661 cgaacaagcc tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa
      721 gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc
      781 tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct
      841 atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca
      901 ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt
      961 cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat
     1021 aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc
     1081 gattgactcg gcagctcatc atgataactc cacaattccg ttggatttta tgcccaggga
     1141 tgctcttcat ggatttgatt ggtctgaaga ggatgacatg tcggatggct tgcccttcct
     1201 gaaaacggac cccaacaata atgggttctt tggcgacggt tctctcttat gtattcttcg
     1261 atctattggc tttaaaccgg aaaattacac gaactctaac gttaacaggc tcccgaccat
     1321 gattacggat agatacacgt tggcttctag atccacaaca tcccgtttac ttcaaagtta
     1381 tctcaataat tttcacccct actgccctat cgtgcactca ccgacgctaa tgatgttgta
     1441 taataaccag attgaaatcg cgtcgaagga tcaatggcaa atccttttta actgcatatt
     1501 agccattgga gcctggtgta tagaggggga atctactgat atagatgttt tttactatca
     1561 aaatgctaaa tctcatttga cgagcaaggt cttcgagtca ggttccataa ttttggtgac
     1621 agccctacat cttctgtcgc gatatacaca gtggaggcag aaaacaaata ctagctataa
     1681 ttttcacagc ttttccataa gaatggccat atcattgggc ttgaataggg acctcccctc
     1741 gtccttcagt gatagcagca ttctggaaca aagacgccga atttggtggt ctgtctactc
     1801 ttgggagatc caattgtccc tgctttatgg tcgatccatc cagctttctc agaatacaat
     1861 ctccttccct tcttctgtcg acgatgtgca gcgtaccaca acaggtccca ccatatatca
     1921 tggcatcatt gaaacagcaa ggctcttaca agttttcaca aaaatctatg aactagacaa
     1981 aacagtaact gcagaaaaaa gtcctatatg tgcaaaaaaa tgcttgatga tttgtaatga
     2041 gattgaggag gtttcgagac aggcaccaaa gtttttacaa atggatattt ccaccaccgc
     2101 tctaaccaat ttgttgaagg aacacccttg gctatccttt acaagattcg aactgaagtg
     2161 gaaacagttg tctcttatca tttatgtatt aagagatttt ttcactaatt ttacccagaa
     2221 aaagtcacaa ctagaacagg atcaaaatga tcatcaaagt tatgaagtta aacgatgctc
     2281 catcatgtta agcgatgcag cacaaagaac tgttatgtct gtaagtagct atatggacaa
     2341 tcataatgtc accccatatt ttgcctggaa ttgttcttat tacttgttca atgcagtcct
     2401 agtacccata aagactctac tctcaaactc aaaatcgaat gctgagaata acgagaccgc
     2461 acaattatta caacaaatta acactgttct gatgctatta aaaaaactgg ccacttttaa
     2521 aatccagact tgtgaaaaat acattcaagt actggaagag gtatgtgcgc cgtttctgtt
     2581 atcacagtgt gcaatcccat taccgcatat cagttataac aatagtaatg gtagcgccat
     2641 taaaaatatc gtcggttctg caactatcgc ccaataccct actcttccgg aggaaaatgt
     2701 caacaatatc agtgttaaat atgtttctcc tggctcagta gggccttcac ctgtgccatt
     2761 gaaatcagga gcaagtttca gtgatctagt caagctgtta tctaaccgtc caccctctcg
     2821 taactctcca gtgacaatac caagaagcac accttcgcat cgctcagtca cgccttttct
     2881 agggcaacag caacagctgc aatcattagt gccactgacc ccgtctgctt tgtttggtgg
     2941 cgccaatttt aatcaaagtg ggaatatcgc tgatagctca ttgtccttca ctttcactaa
     3001 cagtagcaac ggtccgaacc tcataacaac tcaaacaaat tctcaagcgc tttcacaacc
     3061 aattgcctcc tctaacgttc atgataactt catgaataat gaaatcacgg ctagtaaaat
     3121 tgatgatggt aataattcaa aaccactgtc acctggttgg acggaccaaa ctgcgtataa
     3181 cgcgtttgga atcactacag ggatgtttaa taccactaca atggatgatg tatataacta
     3241 tctattcgat gatgaagata ccccaccaaa cccaaaaaaa gaggagggcc gcggcagcct
     3301 gctgacctgc ggcgatgtgg aggagaaccc cgggcccgtg aagatcatct cccgcaagtc
     3361 cctgggcacc cagaacgtgt acgacatcgg cgtggagaag gaccacaact tcctgctgaa
     3421 gaacggcctg gtggcctcca actgcttcaa cgaggtaagt tattgaacaa tggcatcaaa
     3481 tgccttcatc atcactaccc tttagccctt aagaccccac aatgacctta cccactcaga
     3541 gaaaaaagta aatatgaaag cccatttgaa cttctcatca tgatggacca gatgggtgag
     3601 gtggagtacg cgcccgggga gcccaagggc acgccctggc acccgcaccg cggcttcgag
     3661 accgtgacct acatcggcgc gccggtacca ctagtgggga ggttcccttt agtgagggtt
     3721 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3781 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3841 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3901 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3961 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4021 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4081 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4561 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4681 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4741 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4801 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4861 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4921 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4981 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5041 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5101 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5161 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5221 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5281 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5341 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5401 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5461 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5521 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5581 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5641 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5701 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5761 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5821 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5881 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5941 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6001 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6061 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6121 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6181 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6241 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6301 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc
     6361 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6421 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6481 ccagtcacga cg
//
