LOCUS       sequence-436674-        6699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split intein version of Trojan-LexAGAD. Create LexAGAD 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 6699)
  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 6699)
  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)-LexAGAD
FEATURES             Location/Qualifiers
     source          1..6699
                     /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             648..668
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(678..929,930..1283)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     CDS             3492..3545
                     /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    3798..3867
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(3913..3933)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3913..3931)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3952..3968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3952..3968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3965..3987)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3976..3992
                     /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(4000..4006,4007..4024,4025..4030))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4045..4066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4183..4200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4354..4942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4434..4453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5113..5904,5905..5973))
                     /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     5736..5755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5974..6078)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6105..6560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6192..6211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6402..6423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6687..6699,1..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 catgccaccc aagaagaagc
      661 gaaaagtaga agatccaatg aaggctctca cggcccgaca acaggaagtt tttgatttga
      721 tacgggatca tatatcccaa acgggtatgc ctccgacccg cgcagagata gcacagcgac
      781 tgggctttcg atcgcctaac gccgcggagg agcacttgaa ggcactggcc cgcaagggtg
      841 tcattgaaat cgtgtccggt gcgagccgcg gaatccggct gttgcaggag gaggaagagg
      901 gcctgccact ggtgggacgc gtggccgctg gcgagccgct gctggcccag caacacatag
      961 agggacacta tcaggtggac ccctccttgt ttaagccaaa tgctgatttc ctgttgcggg
     1021 tgtcgggaat gtccatgaag gacatcggta ttatggatgg tgacctcctc gccgtccata
     1081 agacacagga tgtcaggaac ggccaggtag tcgttgccag gatagacgat gaggtcactg
     1141 tgaaacgtct caagaagcaa ggcaataagg tcgagctgct gccggagaat agcgagttca
     1201 agccgatcgt ggtggatctg cgacagcagt cctttactat cgagggcttg gccgtgggtg
     1261 tgatccgcaa cggagattgg ctgggatcca tcgacagtgc ggcccaccac gacaacagca
     1321 ccattccact ggacttcatg cctcgtgacg cactgcatgg atttgactgg agcgaggagg
     1381 atgatatgag cgacggactc ccattcctga agaccgaccc caataacaat ggtttctttg
     1441 gcgacggcag cctgctgtgt atcctgcgaa gtattggttt taagccggaa aattacacga
     1501 acagtaatgt caatcgtctt cccacaatga tcaccgaccg ttacacgctg gccagtcgca
     1561 gtactacaag ccgcctgctg caaagctatc ttaacaactt ccacccctac tgccccatag
     1621 tgcatagtcc tactctcatg atgctttaca acaatcagat tgagatagca tcgaaggacc
     1681 agtggcagat cctcttcaac tgcattctcg ccatcggcgc ttggtgtatt gaaggcgaga
     1741 gcacagatat cgacgtgttt tactatcaaa acgccaagtc gcacctcacc tcgaaggtct
     1801 tcgaaagtgg aagcattatc ctcgtgacag ccctccatct gctgagtcgt tatacccagt
     1861 ggcgtcagaa aactaataca tcctacaact ttcattcgtt cagcattcgg atggcgatta
     1921 gcctgggact taatcgtgac ctcccaagta gcttcagcga ttcgtcgatc ctggaacagc
     1981 gccggaggat ttggtggagt gtgtacagct gggagatcca gctgtccctc ttgtacggac
     2041 gcagcattca gctgagtcag aacacgattt ccttcccgtc cagcgtggac gacgttcaac
     2101 ggacaactac gggcccgacc atctaccacg gtatcattga gacggcccgc ttgcttcaag
     2161 tgtttacgaa gatttatgag ctggataaga cagtgaccgc ggagaagagt ccgatctgcg
     2221 ccaagaaatg cctgatgatc tgcaatgaga tagaggaggt gagccgccag gcacccaagt
     2281 tcctgcagat ggacataagc actacggcgc tgactaatct cctcaaagaa cacccctggc
     2341 tgagtttcac ccgcttcgag ctgaagtgga aacagctgag cctgattata tacgttttgc
     2401 gggacttttt cacaaatttt acgcagaaga agtcgcagct tgagcaagat cagaacgacc
     2461 atcagtccta cgaggtaaag aggtgcagca taatgctctc cgacgcagcc cagcgcaccg
     2521 tgatgtcggt gtcgtcctat atggacaacc ataatgtgac cccttatttc gcctggaatt
     2581 gctcctacta tttgttcaac gccgtgttgg tcccaatcaa gactcttctg agcaactcca
     2641 aatcgaacgc cgagaataac gagacagcac agctcttgca gcaaattaac accgtgttga
     2701 tgctgttgaa gaaactcgct acattcaaga ttcagacttg cgaaaagtac atccaggtgt
     2761 tggaagaggt gtgcgcccca ttccttcttt cgcaatgcgc aattcccctc ccgcacataa
     2821 gttacaataa ttccaacggc tccgctatca agaatatcgt cggctccgct acaatcgcgc
     2881 agtatcccac gctgcctgag gaaaatgtta acaacatatc ggtgaagtat gtaagtccag
     2941 gcagcgtcgg cccgagccct gtgccgctga aaagcggagc ctccttcagc gatctggtca
     3001 agctgctcag caaccgccca ccatcccgca actcccctgt cactataccc cgatcgaccc
     3061 cgtcgcatcg tagcgttacc ccattcctgg gacaacagca gcaactgcag agtctggtgc
     3121 cacttacgcc ctcggcattg tttggcggag ccaacttcaa ccagagtggt aacattgcgg
     3181 acagctcctt gagcttcacg tttaccaaca gctccaacgg tccaaacttg atcacgactc
     3241 agacaaactc ccaggcattg tcccagccca ttgccagctc gaatgtgcat gataacttta
     3301 tgaataatga aattactgcc tccaagattg acgacggcaa caacagcaag cccctttccc
     3361 caggctggac tgaccagacg gcctacaatg ctttcggtat caccactggc atgttcaaca
     3421 ccacaacgat ggacgacgta tataactatc tttttgatga tgaggatact ccaccgaacc
     3481 caaagaagga ggaaggacgc ggatcgttgc tgacgtgcgg agacgtggag gagaatccgg
     3541 gccccgtgaa gatcatctcc cgcaagtccc tgggcaccca gaacgtgtac gacatcggcg
     3601 tggagaagga ccacaacttc ctgctgaaga acggcctggt ggcctccaac tgcttcaaca
     3661 ggtaagttat tgaacaatgg catcaaatgc cttcatcatc actacccttt agcccttaag
     3721 accccacaat gaccttaccc actcagagaa aaaagtaaat atgaaagccc atttgaactt
     3781 ctcatcatga tggaccagat gggtgaggtg gagtacgcgc ccggggagcc caagggcacg
     3841 ccctggcacc cgcaccgcgg cttcgagacc gtgacctaca tcggcgcgcc ggtaccacta
     3901 gtggggaggt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     3961 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4021 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4081 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4141 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4201 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4261 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4321 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4381 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4441 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4501 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4561 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4621 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4681 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4741 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4801 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4861 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     6121 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6181 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6241 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6301 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6361 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6421 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6481 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6541 aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6601 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6661 attaagttgg gtaacgccag ggttttccca gtcacgacg
//
