LOCUS       sequence-436675-        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 2 
            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(2)-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             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             642..3284
                     /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             3285..3338
                     /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 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 catgaagcta ctgtcttcta
      661 tcgaacaagc ctgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      721 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      781 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      841 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
      901 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
      961 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1021 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1081 cgattgactc ggcagctcat catgataact ccacaattcc gttggatttt atgcccaggg
     1141 atgctcttca tggatttgat tggtctgaag aggatgacat gtcggatggc ttgcccttcc
     1201 tgaaaacgga ccccaacaat aatgggttct ttggcgacgg ttctctctta tgtattcttc
     1261 gatctattgg ctttaaaccg gaaaattaca cgaactctaa cgttaacagg ctcccgacca
     1321 tgattacgga tagatacacg ttggcttcta gatccacaac atcccgttta cttcaaagtt
     1381 atctcaataa ttttcacccc tactgcccta tcgtgcactc accgacgcta atgatgttgt
     1441 ataataacca gattgaaatc gcgtcgaagg atcaatggca aatccttttt aactgcatat
     1501 tagccattgg agcctggtgt atagaggggg aatctactga tatagatgtt ttttactatc
     1561 aaaatgctaa atctcatttg acgagcaagg tcttcgagtc aggttccata attttggtga
     1621 cagccctaca tcttctgtcg cgatatacac agtggaggca gaaaacaaat actagctata
     1681 attttcacag cttttccata agaatggcca tatcattggg cttgaatagg gacctcccct
     1741 cgtccttcag tgatagcagc attctggaac aaagacgccg aatttggtgg tctgtctact
     1801 cttgggagat ccaattgtcc ctgctttatg gtcgatccat ccagctttct cagaatacaa
     1861 tctccttccc ttcttctgtc gacgatgtgc agcgtaccac aacaggtccc accatatatc
     1921 atggcatcat tgaaacagca aggctcttac aagttttcac aaaaatctat gaactagaca
     1981 aaacagtaac tgcagaaaaa agtcctatat gtgcaaaaaa atgcttgatg atttgtaatg
     2041 agattgagga ggtttcgaga caggcaccaa agtttttaca aatggatatt tccaccaccg
     2101 ctctaaccaa tttgttgaag gaacaccctt ggctatcctt tacaagattc gaactgaagt
     2161 ggaaacagtt gtctcttatc atttatgtat taagagattt tttcactaat tttacccaga
     2221 aaaagtcaca actagaacag gatcaaaatg atcatcaaag ttatgaagtt aaacgatgct
     2281 ccatcatgtt aagcgatgca gcacaaagaa ctgttatgtc tgtaagtagc tatatggaca
     2341 atcataatgt caccccatat tttgcctgga attgttctta ttacttgttc aatgcagtcc
     2401 tagtacccat aaagactcta ctctcaaact caaaatcgaa tgctgagaat aacgagaccg
     2461 cacaattatt acaacaaatt aacactgttc tgatgctatt aaaaaaactg gccactttta
     2521 aaatccagac ttgtgaaaaa tacattcaag tactggaaga ggtatgtgcg ccgtttctgt
     2581 tatcacagtg tgcaatccca ttaccgcata tcagttataa caatagtaat ggtagcgcca
     2641 ttaaaaatat cgtcggttct gcaactatcg cccaataccc tactcttccg gaggaaaatg
     2701 tcaacaatat cagtgttaaa tatgtttctc ctggctcagt agggccttca cctgtgccat
     2761 tgaaatcagg agcaagtttc agtgatctag tcaagctgtt atctaaccgt ccaccctctc
     2821 gtaactctcc agtgacaata ccaagaagca caccttcgca tcgctcagtc acgccttttc
     2881 tagggcaaca gcaacagctg caatcattag tgccactgac cccgtctgct ttgtttggtg
     2941 gcgccaattt taatcaaagt gggaatatcg ctgatagctc attgtccttc actttcacta
     3001 acagtagcaa cggtccgaac ctcataacaa ctcaaacaaa ttctcaagcg ctttcacaac
     3061 caattgcctc ctctaacgtt catgataact tcatgaataa tgaaatcacg gctagtaaaa
     3121 ttgatgatgg taataattca aaaccactgt cacctggttg gacggaccaa actgcgtata
     3181 acgcgtttgg aatcactaca gggatgttta ataccactac aatggatgat gtatataact
     3241 atctattcga tgatgaagat accccaccaa acccaaaaaa agaggaagga cgcggatcgt
     3301 tgctgacgtg cggagacgtg gaggagaatc cgggccccgt gaagatcatc tcccgcaagt
     3361 ccctgggcac ccagaacgtg tacgacatcg gcgtggagaa ggaccacaac ttcctgctga
     3421 agaacggcct ggtggcctcc aactgcttca acaggtaagt 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
//
