LOCUS       sequence-103977-        6599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Creating Gal4 drivers from MiMIC lines with inserts into coding 
            introns in Phase 2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  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 6599)
  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(2)-T2A-Gal4-Hsp70
FEATURES             Location/Qualifiers
     source          1..6599
                     /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             2918..2971
                     /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             2972..5617
                     /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    6303..6372
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(6412..6432)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6412..6430)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6451..6467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6451..6467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6464..6486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6475..6491
                     /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(6499..6505,6506..6523,6524..6529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6544..6565
                     /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 tccgcaggtc
     2881 gggaggttcc ggtggaagcg gaggtagcgg cggatccgag ggccgcggca gcctgctgac
     2941 ctgcggcgat gtggaggaga accccgggcc catgaagctg ctgagcagca tcgagcaggc
     3001 ctgcgatatc tgccgcctga agaagctgaa gtgcagcaag gagaagccca agtgcgccaa
     3061 gtgcctgaag aacaactggg agtgccgcta cagccccaag accaagcgca gccccctgac
     3121 ccgcgcccac ctgaccgagg tggagagccg cctggagcgc ctggagcagc tgttcctgct
     3181 gatcttcccc cgcgaggatc tggatatgat cctgaagatg gatagcctgc aggatatcaa
     3241 ggccctgctg accggcctgt tcgtgcagga taacgtgaac aaggatgccg tgaccgatcg
     3301 cctggccagc gtggagaccg atatgcccct gaccctgcgc cagcaccgca tcagcgccac
     3361 cagcagcagc gaggagagca gcaacaaggg ccagcgccag ctgaccgtga gcatcgatag
     3421 cgccgcccac cacgataaca gcaccatccc cctggatttc atgccccgcg atgccctgca
     3481 cggcttcgat tggagcgagg aggatgatat gagcgatggc ctgcccttcc tgaagaccga
     3541 tcccaacaac aacggcttct tcggcgatgg cagcctgctg tgcatcctgc gcagcatcgg
     3601 cttcaagccc gagaactaca ccaacagcaa cgtgaaccgc ctgcccacca tgatcaccga
     3661 tcgctacacc ctggccagcc gcagcaccac cagccgcctg ctgcagagct acctgaacaa
     3721 cttccacccc tactgcccca tcgtgcacag ccccaccctg atgatgctgt acaacaacca
     3781 gatcgagatc gccagcaagg atcagtggca gatcctgttc aactgcatcc tggccatcgg
     3841 cgcctggtgc atcgagggcg agagcaccga tatcgatgtg ttctactacc agaacgccaa
     3901 gagccacctg accagcaagg tgttcgagag cggcagcatc atcctggtga ccgccctgca
     3961 cctgctgagc cgctacaccc agtggcgcca gaagaccaac accagctaca acttccacag
     4021 cttcagcatc cgcatggcca tcagcctggg cctgaaccgc gatctgccca gcagcttcag
     4081 cgatagcagc atcctggagc agcgccgccg catctggtgg agcgtgtaca gctgggagat
     4141 ccagctgagc ctgctgtacg gccgcagcat ccagctgagc cagaacacca tcagcttccc
     4201 cagcagcgtg gatgatgtgc agcgcaccac caccggcccc accatctacc acggcatcat
     4261 cgagaccgcc cgcctgctgc aggtgttcac caagatctac gagctggata agaccgtgac
     4321 cgccgagaag agccccatct gcgccaagaa gtgcctgatg atctgcaacg agatcgagga
     4381 ggtgagccgc caggccccca agttcctgca gatggatatc agcaccaccg ccctgaccaa
     4441 cctgctgaag gagcacccct ggctgagctt cacccgcttc gagctgaagt ggaagcagct
     4501 gagcctgatc atctacgtgc tgcgcgattt cttcaccaac ttcacccaga agaagagcca
     4561 gctggagcag gatcagaacg atcaccagag ctacgaggtg aagcgctgca gcatcatgct
     4621 gagcgatgcc gcccagcgca ccgtgatgag cgtgagcagc tacatggata accacaacgt
     4681 gaccccctac ttcgcctgga actgcagcta ctacctgttc aacgccgtgc tggtgcccat
     4741 caagaccctg ctgagcaaca gcaagagcaa cgccgagaac aacgagaccg cccagctgct
     4801 gcagcagatc aacaccgtgc tgatgctgct gaagaagctg gccaccttca agatccagac
     4861 ctgcgagaag tacatccagg tgctggagga ggtgtgcgcc cccttcctgc tgagccagtg
     4921 cgccatcccc ctgccccaca tcagctacaa caacagcaac ggcagcgcca tcaagaacat
     4981 cgtgggcagc gccaccatcg cccagtaccc caccctgccc gaggagaacg tgaacaacat
     5041 cagcgtgaag tacgtgagcc ccggcagcgt gggacccagc cccgtgcccc tgaagagcgg
     5101 cgccagcttc agcgatctgg tgaagctgct gagcaaccgc ccccccagcc gcaacagccc
     5161 cgtgaccatc ccccgcagca cccccagcca ccgcagcgtg acccccttcc tgggccagca
     5221 gcagcagctg cagagcctgg tgcccctgac ccccagcgcc ctgttcggcg gcgccaactt
     5281 caaccagagc ggcaacatcg ccgatagcag cctgagcttc accttcacca acagcagcaa
     5341 cggccccaac ctgatcacca cccagaccaa cagccaggcc ctgagccagc ccatcgccag
     5401 cagcaacgtg cacgataact tcatgaacaa cgagatcacc gccagcaaga tcgatgatgg
     5461 caacaacagc aagcccctga gccccggctg gaccgatcag accgcctaca acgccttcgg
     5521 catcaccacc ggcatgttca acaccaccac catggatgat gtgtacaact acctgttcga
     5581 tgatgaggat acccccccca accccaagaa ggagtaaaat gaatcgtaga tactgaaaaa
     5641 ccccgcaagt tcacttcaac tgtgcatcgt gcaccatctc aatttctttc atttatacat
     5701 cgttttgcct tcttttatgt aactatactc ctctaagttt caatcttggc catgtaacct
     5761 ctgatctata gaatttttta aatgactaga attaatgccc atcttttttt tggacctaaa
     5821 ttcttcatga aaatatatta cgagggctta ttcagaagct tatcgatacc gtcgactaaa
     5881 gccaaataga aattattcag ttctggctta agtttttaaa agtgatatta tttatttggt
     5941 tgtaaccaac caaaagaatg taaataacta atacataatt atgttagttt taagttagca
     6001 acaaattgat tttagctata ttagctactt ggttaataaa tagaatatat ttatttaaag
     6061 ataattcgtt tttattgtca gggagtgagt ttgcttaaaa actcgtttgg atccgcggat
     6121 ccggaggtag cggtggaagc ggaggttccg gcaggtaagt tattgaacaa tggcatcaaa
     6181 tgccttcatc atcactaccc tttagccctt aagaccccac aatgacctta cccactcaga
     6241 gaaaaaagta aatatgaaag cccatttgaa cttctaagct tgtcgaccat catgatggac
     6301 cagatgggtg aggtggagta cgcgcccggg gagcccaagg gcacgccctg gcacccgcac
     6361 cgcggcttcg agaccgtgac ctacatcatg catggtaccc agcttttgtt ccctttagtg
     6421 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcgg
//
