LOCUS       sequence-103976-        6591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Creating Gal4 drivers 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 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(1)-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             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             2973..5618
                     /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 gcggaggttc cggtggaagc ggaggtagcg gcggatccga gggccgcggc agcctgctga
     2941 cctgcggcga tgtggaggag aaccccgggc ccatgaagct gctgagcagc atcgagcagg
     3001 cctgcgatat ctgccgcctg aagaagctga agtgcagcaa ggagaagccc aagtgcgcca
     3061 agtgcctgaa gaacaactgg gagtgccgct acagccccaa gaccaagcgc agccccctga
     3121 cccgcgccca cctgaccgag gtggagagcc gcctggagcg cctggagcag ctgttcctgc
     3181 tgatcttccc ccgcgaggat ctggatatga tcctgaagat ggatagcctg caggatatca
     3241 aggccctgct gaccggcctg ttcgtgcagg ataacgtgaa caaggatgcc gtgaccgatc
     3301 gcctggccag cgtggagacc gatatgcccc tgaccctgcg ccagcaccgc atcagcgcca
     3361 ccagcagcag cgaggagagc agcaacaagg gccagcgcca gctgaccgtg agcatcgata
     3421 gcgccgccca ccacgataac agcaccatcc ccctggattt catgccccgc gatgccctgc
     3481 acggcttcga ttggagcgag gaggatgata tgagcgatgg cctgcccttc ctgaagaccg
     3541 atcccaacaa caacggcttc ttcggcgatg gcagcctgct gtgcatcctg cgcagcatcg
     3601 gcttcaagcc cgagaactac accaacagca acgtgaaccg cctgcccacc atgatcaccg
     3661 atcgctacac cctggccagc cgcagcacca ccagccgcct gctgcagagc tacctgaaca
     3721 acttccaccc ctactgcccc atcgtgcaca gccccaccct gatgatgctg tacaacaacc
     3781 agatcgagat cgccagcaag gatcagtggc agatcctgtt caactgcatc ctggccatcg
     3841 gcgcctggtg catcgagggc gagagcaccg atatcgatgt gttctactac cagaacgcca
     3901 agagccacct gaccagcaag gtgttcgaga gcggcagcat catcctggtg accgccctgc
     3961 acctgctgag ccgctacacc cagtggcgcc agaagaccaa caccagctac aacttccaca
     4021 gcttcagcat ccgcatggcc atcagcctgg gcctgaaccg cgatctgccc agcagcttca
     4081 gcgatagcag catcctggag cagcgccgcc gcatctggtg gagcgtgtac agctgggaga
     4141 tccagctgag cctgctgtac ggccgcagca tccagctgag ccagaacacc atcagcttcc
     4201 ccagcagcgt ggatgatgtg cagcgcacca ccaccggccc caccatctac cacggcatca
     4261 tcgagaccgc ccgcctgctg caggtgttca ccaagatcta cgagctggat aagaccgtga
     4321 ccgccgagaa gagccccatc tgcgccaaga agtgcctgat gatctgcaac gagatcgagg
     4381 aggtgagccg ccaggccccc aagttcctgc agatggatat cagcaccacc gccctgacca
     4441 acctgctgaa ggagcacccc tggctgagct tcacccgctt cgagctgaag tggaagcagc
     4501 tgagcctgat catctacgtg ctgcgcgatt tcttcaccaa cttcacccag aagaagagcc
     4561 agctggagca ggatcagaac gatcaccaga gctacgaggt gaagcgctgc agcatcatgc
     4621 tgagcgatgc cgcccagcgc accgtgatga gcgtgagcag ctacatggat aaccacaacg
     4681 tgacccccta cttcgcctgg aactgcagct actacctgtt caacgccgtg ctggtgccca
     4741 tcaagaccct gctgagcaac agcaagagca acgccgagaa caacgagacc gcccagctgc
     4801 tgcagcagat caacaccgtg ctgatgctgc tgaagaagct ggccaccttc aagatccaga
     4861 cctgcgagaa gtacatccag gtgctggagg aggtgtgcgc ccccttcctg ctgagccagt
     4921 gcgccatccc cctgccccac atcagctaca acaacagcaa cggcagcgcc atcaagaaca
     4981 tcgtgggcag cgccaccatc gcccagtacc ccaccctgcc cgaggagaac gtgaacaaca
     5041 tcagcgtgaa gtacgtgagc cccggcagcg tgggacccag ccccgtgccc ctgaagagcg
     5101 gcgccagctt cagcgatctg gtgaagctgc tgagcaaccg cccccccagc cgcaacagcc
     5161 ccgtgaccat cccccgcagc acccccagcc accgcagcgt gacccccttc ctgggccagc
     5221 agcagcagct gcagagcctg gtgcccctga cccccagcgc cctgttcggc ggcgccaact
     5281 tcaaccagag cggcaacatc gccgatagca gcctgagctt caccttcacc aacagcagca
     5341 acggccccaa cctgatcacc acccagacca acagccaggc cctgagccag cccatcgcca
     5401 gcagcaacgt gcacgataac ttcatgaaca acgagatcac cgccagcaag atcgatgatg
     5461 gcaacaacag caagcccctg agccccggct ggaccgatca gaccgcctac aacgccttcg
     5521 gcatcaccac cggcatgttc aacaccacca ccatggatga tgtgtacaac tacctgttcg
     5581 atgatgagga tacccccccc aaccccaaga aggagtaaaa tgaatcgtag atactgaaaa
     5641 accccgcaag ttcacttcaa ctgtgcatcg tgcaccatct caatttcttt catttataca
     5701 tcgttttgcc ttcttttatg taactatact cctctaagtt tcaatcttgg ccatgtaacc
     5761 tctgatctat agaatttttt aaatgactag aattaatgcc catctttttt ttggacctaa
     5821 attcttcatg aaaatatatt acgagggctt attcagaagc ttatcgatac cgtcgactaa
     5881 agccaaatag aaattattca gttctggctt aagtttttaa aagtgatatt atttatttgg
     5941 ttgtaaccaa ccaaaagaat gtaaataact aatacataat tatgttagtt ttaagttagc
     6001 aacaaattga ttttagctat attagctact tggttaataa atagaatata tttatttaaa
     6061 gataattcgt ttttattgtc agggagtgag tttgcttaaa aactcgtttg gatccggagg
     6121 tagcggtgga agcggaggtt ccggaggtaa 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
//
