LOCUS       sequence-103975-        6591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Creating Gal4 drivers from MiMIC lines with inserts into coding 
            introns in Phase 0..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6591)
  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 6591)
  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(0)-T2A-Gal4-Hsp70
FEATURES             Location/Qualifiers
     source          1..6591
                     /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             2917..2970
                     /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             2971..5616
                     /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"
     protein_bind    6295..6364
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(6404..6424)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6404..6422)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6443..6459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6443..6459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6456..6478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6467..6483
                     /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(6491..6497,6498..6515,6516..6521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6536..6557
                     /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 ggaggttccg gtggaagcgg aggtagcggc ggatccgagg gccgcggcag cctgctgacc
     2941 tgcggcgatg tggaggagaa ccccgggccc atgaagctgc tgagcagcat cgagcaggcc
     3001 tgcgatatct gccgcctgaa gaagctgaag tgcagcaagg agaagcccaa gtgcgccaag
     3061 tgcctgaaga acaactggga gtgccgctac agccccaaga ccaagcgcag ccccctgacc
     3121 cgcgcccacc tgaccgaggt ggagagccgc ctggagcgcc tggagcagct gttcctgctg
     3181 atcttccccc gcgaggatct ggatatgatc ctgaagatgg atagcctgca ggatatcaag
     3241 gccctgctga ccggcctgtt cgtgcaggat aacgtgaaca aggatgccgt gaccgatcgc
     3301 ctggccagcg tggagaccga tatgcccctg accctgcgcc agcaccgcat cagcgccacc
     3361 agcagcagcg aggagagcag caacaagggc cagcgccagc tgaccgtgag catcgatagc
     3421 gccgcccacc acgataacag caccatcccc ctggatttca tgccccgcga tgccctgcac
     3481 ggcttcgatt ggagcgagga ggatgatatg agcgatggcc tgcccttcct gaagaccgat
     3541 cccaacaaca acggcttctt cggcgatggc agcctgctgt gcatcctgcg cagcatcggc
     3601 ttcaagcccg agaactacac caacagcaac gtgaaccgcc tgcccaccat gatcaccgat
     3661 cgctacaccc tggccagccg cagcaccacc agccgcctgc tgcagagcta cctgaacaac
     3721 ttccacccct actgccccat cgtgcacagc cccaccctga tgatgctgta caacaaccag
     3781 atcgagatcg ccagcaagga tcagtggcag atcctgttca actgcatcct ggccatcggc
     3841 gcctggtgca tcgagggcga gagcaccgat atcgatgtgt tctactacca gaacgccaag
     3901 agccacctga ccagcaaggt gttcgagagc ggcagcatca tcctggtgac cgccctgcac
     3961 ctgctgagcc gctacaccca gtggcgccag aagaccaaca ccagctacaa cttccacagc
     4021 ttcagcatcc gcatggccat cagcctgggc ctgaaccgcg atctgcccag cagcttcagc
     4081 gatagcagca tcctggagca gcgccgccgc atctggtgga gcgtgtacag ctgggagatc
     4141 cagctgagcc tgctgtacgg ccgcagcatc cagctgagcc agaacaccat cagcttcccc
     4201 agcagcgtgg atgatgtgca gcgcaccacc accggcccca ccatctacca cggcatcatc
     4261 gagaccgccc gcctgctgca ggtgttcacc aagatctacg agctggataa gaccgtgacc
     4321 gccgagaaga gccccatctg cgccaagaag tgcctgatga tctgcaacga gatcgaggag
     4381 gtgagccgcc aggcccccaa gttcctgcag atggatatca gcaccaccgc cctgaccaac
     4441 ctgctgaagg agcacccctg gctgagcttc acccgcttcg agctgaagtg gaagcagctg
     4501 agcctgatca tctacgtgct gcgcgatttc ttcaccaact tcacccagaa gaagagccag
     4561 ctggagcagg atcagaacga tcaccagagc tacgaggtga agcgctgcag catcatgctg
     4621 agcgatgccg cccagcgcac cgtgatgagc gtgagcagct acatggataa ccacaacgtg
     4681 accccctact tcgcctggaa ctgcagctac tacctgttca acgccgtgct ggtgcccatc
     4741 aagaccctgc tgagcaacag caagagcaac gccgagaaca acgagaccgc ccagctgctg
     4801 cagcagatca acaccgtgct gatgctgctg aagaagctgg ccaccttcaa gatccagacc
     4861 tgcgagaagt acatccaggt gctggaggag gtgtgcgccc ccttcctgct gagccagtgc
     4921 gccatccccc tgccccacat cagctacaac aacagcaacg gcagcgccat caagaacatc
     4981 gtgggcagcg ccaccatcgc ccagtacccc accctgcccg aggagaacgt gaacaacatc
     5041 agcgtgaagt acgtgagccc cggcagcgtg ggacccagcc ccgtgcccct gaagagcggc
     5101 gccagcttca gcgatctggt gaagctgctg agcaaccgcc cccccagccg caacagcccc
     5161 gtgaccatcc cccgcagcac ccccagccac cgcagcgtga cccccttcct gggccagcag
     5221 cagcagctgc agagcctggt gcccctgacc cccagcgccc tgttcggcgg cgccaacttc
     5281 aaccagagcg gcaacatcgc cgatagcagc ctgagcttca ccttcaccaa cagcagcaac
     5341 ggccccaacc tgatcaccac ccagaccaac agccaggccc tgagccagcc catcgccagc
     5401 agcaacgtgc acgataactt catgaacaac gagatcaccg ccagcaagat cgatgatggc
     5461 aacaacagca agcccctgag ccccggctgg accgatcaga ccgcctacaa cgccttcggc
     5521 atcaccaccg gcatgttcaa caccaccacc atggatgatg tgtacaacta cctgttcgat
     5581 gatgaggata ccccccccaa ccccaagaag gagtaaaatg aatcgtagat actgaaaaac
     5641 cccgcaagtt cacttcaact gtgcatcgtg caccatctca atttctttca tttatacatc
     5701 gttttgcctt cttttatgta actatactcc tctaagtttc aatcttggcc atgtaacctc
     5761 tgatctatag aattttttaa atgactagaa ttaatgccca tctttttttt ggacctaaat
     5821 tcttcatgaa aatatattac gagggcttat tcagaagctt atcgataccg tcgactaaag
     5881 ccaaatagaa attattcagt tctggcttaa gtttttaaaa gtgatattat ttatttggtt
     5941 gtaaccaacc aaaagaatgt aaataactaa tacataatta tgttagtttt aagttagcaa
     6001 caaattgatt ttagctatat tagctacttg gttaataaat agaatatatt tatttaaaga
     6061 taattcgttt ttattgtcag ggagtgagtt tgcttaaaaa ctcgtttgga tccggaggta
     6121 gcggtggaag cggaggttcc ggcgaggtaa gttattgaac aatggcatca aatgccttca
     6181 tcatcactac cctttagccc ttaagacccc acaatgacct tacccactca gagaaaaaag
     6241 taaatatgaa agcccatttg aacttctaag cttgtcgacc atcatgatgg accagatggg
     6301 tgaggtggag tacgcgcccg gggagcccaa gggcacgccc tggcacccgc accgcggctt
     6361 cgagaccgtg acctacatca tgcatggtac ccagcttttg ttccctttag tgagggttaa
     6421 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6481 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6541 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg g
//
