LOCUS       sequence-313664-        6579 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 6579)
  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 6579)
  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
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..54
                     /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             55..2700
                     /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    2889..2958
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(2998..3016)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3037..3053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3061..3077
                     /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(3085..3091,3092..3109,3110..3115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3130..3151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    3365..3434
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(3474..3492)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3513..3529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3537..3553
                     /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(3561..3567,3568..3585,3586..3591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3606..3627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3744..3761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3915..4503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3995..4014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4674..5465,5466..5534))
                     /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     5297..5316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5535..5639)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5665..6120)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5802..5823)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6014..6033
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6247..6269
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6261..6278
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6262..6278
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6288..6307
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6288..6306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6342..6411
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 gagggccgcg gcagcctgct gacctgcggc gatgtggagg agaaccccgg gcccatgaag
       61 ctgctgagca gcatcgagca ggcctgcgat atctgccgcc tgaagaagct gaagtgcagc
      121 aaggagaagc ccaagtgcgc caagtgcctg aagaacaact gggagtgccg ctacagcccc
      181 aagaccaagc gcagccccct gacccgcgcc cacctgaccg aggtggagag ccgcctggag
      241 cgcctggagc agctgttcct gctgatcttc ccccgcgagg atctggatat gatcctgaag
      301 atggatagcc tgcaggatat caaggccctg ctgaccggcc tgttcgtgca ggataacgtg
      361 aacaaggatg ccgtgaccga tcgcctggcc agcgtggaga ccgatatgcc cctgaccctg
      421 cgccagcacc gcatcagcgc caccagcagc agcgaggaga gcagcaacaa gggccagcgc
      481 cagctgaccg tgagcatcga tagcgccgcc caccacgata acagcaccat ccccctggat
      541 ttcatgcccc gcgatgccct gcacggcttc gattggagcg aggaggatga tatgagcgat
      601 ggcctgccct tcctgaagac cgatcccaac aacaacggct tcttcggcga tggcagcctg
      661 ctgtgcatcc tgcgcagcat cggcttcaag cccgagaact acaccaacag caacgtgaac
      721 cgcctgccca ccatgatcac cgatcgctac accctggcca gccgcagcac caccagccgc
      781 ctgctgcaga gctacctgaa caacttccac ccctactgcc ccatcgtgca cagccccacc
      841 ctgatgatgc tgtacaacaa ccagatcgag atcgccagca aggatcagtg gcagatcctg
      901 ttcaactgca tcctggccat cggcgcctgg tgcatcgagg gcgagagcac cgatatcgat
      961 gtgttctact accagaacgc caagagccac ctgaccagca aggtgttcga gagcggcagc
     1021 atcatcctgg tgaccgccct gcacctgctg agccgctaca cccagtggcg ccagaagacc
     1081 aacaccagct acaacttcca cagcttcagc atccgcatgg ccatcagcct gggcctgaac
     1141 cgcgatctgc ccagcagctt cagcgatagc agcatcctgg agcagcgccg ccgcatctgg
     1201 tggagcgtgt acagctggga gatccagctg agcctgctgt acggccgcag catccagctg
     1261 agccagaaca ccatcagctt ccccagcagc gtggatgatg tgcagcgcac caccaccggc
     1321 cccaccatct accacggcat catcgagacc gcccgcctgc tgcaggtgtt caccaagatc
     1381 tacgagctgg ataagaccgt gaccgccgag aagagcccca tctgcgccaa gaagtgcctg
     1441 atgatctgca acgagatcga ggaggtgagc cgccaggccc ccaagttcct gcagatggat
     1501 atcagcacca ccgccctgac caacctgctg aaggagcacc cctggctgag cttcacccgc
     1561 ttcgagctga agtggaagca gctgagcctg atcatctacg tgctgcgcga tttcttcacc
     1621 aacttcaccc agaagaagag ccagctggag caggatcaga acgatcacca gagctacgag
     1681 gtgaagcgct gcagcatcat gctgagcgat gccgcccagc gcaccgtgat gagcgtgagc
     1741 agctacatgg ataaccacaa cgtgaccccc tacttcgcct ggaactgcag ctactacctg
     1801 ttcaacgccg tgctggtgcc catcaagacc ctgctgagca acagcaagag caacgccgag
     1861 aacaacgaga ccgcccagct gctgcagcag atcaacaccg tgctgatgct gctgaagaag
     1921 ctggccacct tcaagatcca gacctgcgag aagtacatcc aggtgctgga ggaggtgtgc
     1981 gcccccttcc tgctgagcca gtgcgccatc cccctgcccc acatcagcta caacaacagc
     2041 aacggcagcg ccatcaagaa catcgtgggc agcgccacca tcgcccagta ccccaccctg
     2101 cccgaggaga acgtgaacaa catcagcgtg aagtacgtga gccccggcag cgtgggaccc
     2161 agccccgtgc ccctgaagag cggcgccagc ttcagcgatc tggtgaagct gctgagcaac
     2221 cgccccccca gccgcaacag ccccgtgacc atcccccgca gcacccccag ccaccgcagc
     2281 gtgaccccct tcctgggcca gcagcagcag ctgcagagcc tggtgcccct gacccccagc
     2341 gccctgttcg gcggcgccaa cttcaaccag agcggcaaca tcgccgatag cagcctgagc
     2401 ttcaccttca ccaacagcag caacggcccc aacctgatca ccacccagac caacagccag
     2461 gccctgagcc agcccatcgc cagcagcaac gtgcacgata acttcatgaa caacgagatc
     2521 accgccagca agatcgatga tggcaacaac agcaagcccc tgagccccgg ctggaccgat
     2581 cagaccgcct acaacgcctt cggcatcacc accggcatgt tcaacaccac caccatggat
     2641 gatgtgtaca actacctgtt cgatgatgag gatacccccc ccaaccccaa gaaggagtaa
     2701 taaggatccg gaggtagcgg tggaagcgga ggttccggag gtaagttatt gaacaatggc
     2761 atcaaatgcc ttcatcatca ctacccttta gcccttaaga ccccacaatg accttaccca
     2821 ctcagagaaa aaagtaaata tgaaagccca tttgaacttc taagcttgtc gaccatcatg
     2881 atggaccaga tgggtgaggt ggagtacgcg cccggggagc ccaagggcac gccctggcac
     2941 ccgcaccgcg gcttcgagac cgtgacctac atcatgcatg gtacccagct tttgttccct
     3001 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3061 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     3121 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3181 gtcggggagg tagcggtgga agcggaggtt ccggaggtaa gttattgaac aatggcatca
     3241 aatgccttca tcatcactac cctttagccc ttaagacccc acaatgacct tacccactca
     3301 gagaaaaaag taaatatgaa agcccatttg aacttctaag cttgtcgacc atcatgatgg
     3361 accagatggg tgaggtggag tacgcgcccg gggagcccaa gggcacgccc tggcacccgc
     3421 accgcggctt cgagaccgtg acctacatca tgcatggtac ccagcttttg ttccctttag
     3481 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3541 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     3601 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3661 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3721 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4081 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4201 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4261 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4321 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4381 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4441 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4501 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4561 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4621 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4921 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4981 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5041 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5101 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5161 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5221 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5281 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5341 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5401 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5461 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5521 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5581 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5641 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     5701 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5761 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5821 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5881 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5941 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6001 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6061 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6121 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6181 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     6241 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     6301 tatagggcga attggagctc gctagcgatg taggtcacgg tctcgaagcc gcggtgcggg
     6361 tgccagggcg tgcccttggg ctccccgggc gcgtactcca cctcacccat ctggtccatc
     6421 atgatggtcg actctagaag tcgatccaac atggcgactt gtcccatccc cggcatgttt
     6481 aaatatacta attattcttg aactaatttt aatcaaccga tttatctctc ttccgcaggt
     6541 ggcggaggtt ccggtggaag cggaggtagc ggcggatcc
//
