LOCUS       sequence-146180-        6823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Creating LexA::GADfluw lines from MiMIC lines with inserts into 
            coding introns in Phase 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  AUTHORS   Diao F, Ironfield H, Luan H, Diao F, Shropshire WC, Ewer J, Marr E, 
            Potter CJ, Landgraf M, White BH
  TITLE     Plug-and-play genetic access to drosophila cell types using 
            exchangeable exon cassettes.
  JOURNAL   Cell Rep. 2015 Mar 3;10(8):1410-21. doi: 
            10.1016/j.celrep.2015.01.059. Epub 2015 Feb 26.
  PUBMED    25732830
REFERENCE   2  (bases 1 to 6823)
  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"; doi: 
            "10.1016/j.celrep.2015.01.059"; journalName: "Cell Rep"; date: 
            "2015-03-3- 3"; volume: "10"; issue: "8"; pages: "1410-21"
COMMENT     Sequence Label: pBS-KS-attB2-SA(1)-T2A-LexA::GADfluw-Hsp70
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(83..100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(254..842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(334..353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1013..1804,1805..1873))
                     /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     1636..1655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1874..1978)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2004..2459)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2141..2162)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2353..2372
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2586..2608
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2600..2617
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2601..2617
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2627..2646
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2627..2645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2681..2750
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             2919..2972
                     /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             2979..2999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3009..3260,3261..3614)
                     /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"
     protein_bind    6527..6596
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(6636..6656)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6636..6654)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6675..6691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6675..6691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6688..6710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6699..6715
                     /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(6723..6729,6730..6747,6748..6753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6768..6789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
       61 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      661 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
      721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1021 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1081 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1141 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1201 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1261 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1321 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1381 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1441 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1501 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1561 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1621 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1681 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1741 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1801 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1861 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1921 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1981 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     2041 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2101 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2161 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2221 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2281 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2341 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2401 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2461 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2521 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     2581 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     2641 atagggcgaa ttggagctcg ctagcgatgt aggtcacggt ctcgaagccg cggtgcgggt
     2701 gccagggcgt gcccttgggc tccccgggcg cgtactccac ctcacccatc tggtccatca
     2761 tgatggtcga ctctagaagt cgatccaaca tggcgacttg tcccatcccc ggcatgttta
     2821 aatatactaa ttattcttga actaatttta atcaaccgat ttatctctct tccgcaggtg
     2881 gcggaggttc cggtggaagc ggaggtagcg gcggatccga gggccgcggc agcctgctga
     2941 cctgcggcga tgtggaggag aaccccgggc ccatgccacc caagaagaag cgaaaagtag
     3001 aagatccaat gaaggctctc acggcccgac aacaggaagt ttttgatttg atacgggatc
     3061 atatatccca aacgggtatg cctccgaccc gcgcagagat agcacagcga ctgggctttc
     3121 gatcgcctaa cgccgcggag gagcacttga aggcactggc ccgcaagggt gtcattgaaa
     3181 tcgtgtccgg tgcgagccgc ggaatccggc tgttgcagga ggaggaagag ggcctgccac
     3241 tggtgggacg cgtggccgct ggcgagccgc tgctggccca gcaacacata gagggacact
     3301 atcaggtgga cccctccttg tttaagccaa atgctgattt cctgttgcgg gtgtcgggaa
     3361 tgtccatgaa ggacatcggt attatggatg gtgacctcct cgccgtccat aagacacagg
     3421 atgtcaggaa cggccaggta gtcgttgcca ggatagacga tgaggtcact gtgaaacgtc
     3481 tcaagaagca aggcaataag gtcgagctgc tgccggagaa tagcgagttc aagccgatcg
     3541 tggtggatct gcgacagcag tcctttacta tcgagggctt ggccgtgggt gtgatccgca
     3601 acggagattg gctgggatcc atcgacagtg cggcccacca cgacaacagc accattccac
     3661 tggacttcat gcctcgtgac gcactgcatg gatttgactg gagcgaggag gatgatatga
     3721 gcgacggact cccattcctg aagaccgacc ccaataacaa tggtttcttt ggcgacggca
     3781 gcctgctgtg tatcctgcga agtattggtt ttaagccgga aaattacacg aacagtaatg
     3841 tcaatcgtct tcccacaatg atcaccgacc gttacacgct ggccagtcgc agtactacaa
     3901 gccgcctgct gcaaagctat cttaacaact tccaccccta ctgccccata gtgcatagtc
     3961 ctactctcat gatgctttac aacaatcaga ttgagatagc atcgaaggac cagtggcaga
     4021 tcctcttcaa ctgcattctc gccatcggcg cttggtgtat tgaaggcgag agcacagata
     4081 tcgacgtgtt ttactatcaa aacgccaagt cgcacctcac ctcgaaggtc ttcgaaagtg
     4141 gaagcattat cctcgtgaca gccctccatc tgctgagtcg ttatacccag tggcgtcaga
     4201 aaactaatac atcctacaac tttcattcgt tcagcattcg gatggcgatt agcctgggac
     4261 ttaatcgtga cctcccaagt agcttcagcg attcgtcgat cctggaacag cgccggagga
     4321 tttggtggag tgtgtacagc tgggagatcc agctgtccct cttgtacgga cgcagcattc
     4381 agctgagtca gaacacgatt tccttcccgt ccagcgtgga cgacgttcaa cggacaacta
     4441 cgggcccgac catctaccac ggtatcattg agacggcccg cttgcttcaa gtgtttacga
     4501 agatttatga gctggataag acagtgaccg cggagaagag tccgatctgc gccaagaaat
     4561 gcctgatgat ctgcaatgag atagaggagg tgagccgcca ggcacccaag ttcctgcaga
     4621 tggacataag cactacggcg ctgactaatc tcctcaaaga acacccctgg ctgagtttca
     4681 cccgcttcga gctgaagtgg aaacagctga gtctgattat atacgttttg cgggactttt
     4741 tcacaaattt tacgcagaag aagtcgcagc ttgagcaaga tcagaacgac catcagtcct
     4801 acgaggtaaa gaggtgcagc ataatgctct ccgacgcagc ccagcgcacc gtgatgtcgg
     4861 tgtcgtccta tatggacaac cataatgtga ccccttattt cgcctggaat tgctcctact
     4921 atttgttcaa cgccgtgttg gtcccaatca agactcttct gagcaactcc aaatcgaacg
     4981 ccgagaataa cgagacagca cagctcttgc agcaaattaa caccgtgttg atgctgttga
     5041 agaaactcgc tacattcaag attcagactt gcgaaaagta catccaggtg ttggaagagg
     5101 tgtgcgcccc attccttctt tcgcaatgcg caattcccct cccgcacata agttacaata
     5161 attccaacgg ctccgctatc aagaatatcg tcggctccgc tacaatcgcg cagtatccca
     5221 cgctgcctga ggaaaatgtt aacaacatat cggtgaagta tgtaagtcca ggcagcgtcg
     5281 gcccgagccc tgtgccgctg aaaagcggag cctccttcag cgatctggtc aagctgctca
     5341 gcaaccgccc accatcccgc aactcccctg tcactatacc ccgatcgacc ccgtcgcatc
     5401 gtagcgttac cccattcctg ggacaacagc agcaactgca gagtctggtg ccacttacgc
     5461 cctcggcatt gtttggcgga gccaacttca accagagtgg taacattgcg gacagctcct
     5521 tgagcttcac gtttaccaac agctccaacg gtccaaactt gatcacgact cagacaaact
     5581 cccaggcatt gtcccagccc attgccagct cgaatgtgca tgataacttt atgaataatg
     5641 aaattactgc ctccaagatt gacgacggca acaacagcaa gcccctttcc ccaggctgga
     5701 ctgaccagac ggcctacaat gctttcggta tcaccactgg catgttcaac accacaacga
     5761 tggacgacgt atataactat ctttttgatg atgaggatac tccaccgaac ccaaagaagg
     5821 agtagcctag gttaattaat aaaatgaatc gtagatactg aaaaaccccg caagttcact
     5881 tcaactgtgc atcgtgcacc atctcaattt ctttcattta tacatcgttt tgccttcttt
     5941 tatgtaacta tactcctcta agtttcaatc ttggccatgt aacctctgat ctatagaatt
     6001 ttttaaatga ctagaattaa tgcccatctt ttttttggac ctaaattctt catgaaaata
     6061 tattacgagg gcttattcag aagcttatcg ataccgtcga ctaaagccaa atagaaatta
     6121 ttcagttctg gcttaagttt ttaaaagtga tattatttat ttggttgtaa ccaaccaaaa
     6181 gaatgtaaat aactaataca taattatgtt agttttaagt tagcaacaaa ttgattttag
     6241 ctatattagc tacttggtta ataaatagaa tatatttatt taaagataat tcgtttttat
     6301 tgtcagggag tgagtttgct taaaaactcg tttttaatta aagatccgga ggtagcggtg
     6361 gaagcggagg ttccggaggt aagttattga acaatggcat caaatgcctt catcatcact
     6421 accctttagc ccttaagacc ccacaatgac cttacccact cagagaaaaa agtaaatatg
     6481 aaagcccatt tgaacttcta agcttgtcga ccatcatgat ggaccagatg ggtgaggtgg
     6541 agtacgcgcc cggggagccc aagggcacgc cctggcaccc gcaccgcggc ttcgagaccg
     6601 tgacctacat catgcatggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     6661 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgg
//
