LOCUS       sequence-104092-        8748 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Creating Gal80 lines from MiMIC lines with inserts into coding 
            introns in Phase 1. Gal80 is tagged with FLAG, HA and EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8748)
  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 8748)
  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-3X Gal80-Hsp70
FEATURES             Location/Qualifiers
     source          1..8748
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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             4277..4303
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4307..4330
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4331..4387
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5693..6406
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             6407..6460
                     /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"
     protein_bind    8452..8521
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(8561..8581)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8561..8579)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8600..8616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8600..8616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8613..8635)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8624..8640
                     /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(8648..8654,8655..8672,8673..8678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8693..8714
                     /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 catggattac aacaaaagga gtagtgtgag
     3001 tacggtgccg aatgctgctc ccattcgcgt gggcttcgtg ggattgaacg cggctaaggg
     3061 ttgggccatt aaaacgcatt atccagccat actgcagctg agctcccagt tccaaataac
     3121 agcgctttat tcccccaaga tcgagacgtc cattgcgaca attcagaggc tgaagttgtc
     3181 caacgccaca gctttcccaa cactcgagag cttcgcctcg agctccacga tcgatatgat
     3241 tgtgatcgca atccaggtgg cttcccacta cgaggtggta atgcccctgc tcgagttttc
     3301 gaagaacaat cctaatctga agtatttgtt tgtggaatgg gcactggcct gctcgctcga
     3361 tcaggcggag tcgatctata aggccgctgc agagcggggc gtgcaaacaa taatcagtct
     3421 gcagggacga aagagcccgt acattttgcg ggcaaaagag cttatctccc agggctacat
     3481 cggcgatata aatagcatcg agattgcagg taacggaggt tggtacggtt acgagcggcc
     3541 ggtgaaaagc ccgaaataca tttacgagat cggaaacgga gttgatctgg tgaccaccac
     3601 gttcggtcac acgatagata tattgcagta catgaccagt tcgtacttca gccgtatcaa
     3661 tgccatggtg tttaacaaca ttccagaaca ggagctcatt gatgagaggg gcaatcgcct
     3721 gggccagcgg gtcccaaaga ctgtcccaga tcatctcttg ttccagggca cgctgctcaa
     3781 cggaaatgtt cctgtgtcct gttcctttaa gggaggcaag cctacgaaaa agttcaccaa
     3841 gaatctggtc atagatatcc atggtacaaa gggtgatctg aagctggagg gtgacgctgg
     3901 atttgcggaa atctcgaatc tcgttctgta ttattcgggc acccgcgcca atgattttcc
     3961 cctggccaat ggtcagcagg cacccttgga ccccggctac gacgcaggca aggagattat
     4021 ggaggtgtat cacctccgaa actacaacgc gatcgttggt aatatacacc ggctctacca
     4081 gagtatcagc gatttccact tcaacaccaa gaaaatcccc gagcttccat cgcagttcgt
     4141 aatgcaaggc ttcgatttcg aaggctttcc caccctgatg gatgccttga tcttgcatcg
     4201 cctgattgaa agtgtctaca agtcgaatat gatgggcagt accctgaatg tttccaacat
     4261 ctcccactac tcgctttacc cctacgatgt gcccgattac gccggcgatt acaaggacga
     4321 tgacgacaag gccaccaact tcagcctgct gaagcaggcc ggcgatgtgg aggagaaccc
     4381 cgggcccatg gattacaaca aaaggagtag tgtgagtacg gtgccgaatg ctgctcccat
     4441 tcgcgtgggc ttcgtgggat tgaacgcggc taagggttgg gccattaaaa cgcattatcc
     4501 agccatactg cagctgagct cccagttcca aataacagcg ctttattccc ccaagatcga
     4561 gacgtccatt gcgacaattc agaggctgaa gttgtccaac gccacagctt tcccaacact
     4621 cgagagcttc gcctcgagct ccacgatcga tatgattgtg atcgcaatcc aggtggcttc
     4681 ccactacgag gtggtaatgc ccctgctcga gttttcgaag aacaatccta atctgaagta
     4741 tttgtttgtg gaatgggcac tggcctgctc gctcgatcag gcggagtcga tctataaggc
     4801 cgctgcagag cggggcgtgc aaacaataat cagtctgcag ggacgaaaga gcccgtacat
     4861 tttgcgggca aaagagctta tctcccaggg ctacatcggc gatataaata gcatcgagat
     4921 tgcaggtaac ggaggttggt acggttacga gcggccggtg aaaagcccga aatacattta
     4981 cgagatcgga aacggagttg atctggtgac caccacgttc ggtcacacga tagatatatt
     5041 gcagtacatg accagttcgt acttcagccg tatcaatgcc atggtgttta acaacattcc
     5101 agaacaggag ctcattgatg agaggggcaa tcgcctgggc cagcgggtcc caaagactgt
     5161 cccagatcat ctcttgttcc agggcacgct gctcaacgga aatgttcctg tgtcctgttc
     5221 ctttaaggga ggcaagccta cgaaaaagtt caccaagaat ctggtcatag atatccatgg
     5281 tacaaagggt gatctgaagc tggagggtga cgctggattt gcggaaatct cgaatctcgt
     5341 tctgtattat tcgggcaccc gcgccaatga ttttcccctg gccaatggtc agcaggcacc
     5401 cttggacccc ggctacgacg caggcaagga gattatggag gtgtatcacc tccgaaacta
     5461 caacgcgatc gttggtaata tacaccggct ctaccagagt atcagcgatt tccacttcaa
     5521 caccaagaaa atccccgagc ttccatcgca gttcgtaatg caaggcttcg atttcgaagg
     5581 ctttcccacc ctgatggatg ccttgatctt gcatcgcctg attgaaagtg tctacaagtc
     5641 gaatatgatg ggcagtaccc tgaatgtttc caacatctcc cactactcgc ttatgtccaa
     5701 aggtgaagaa ctgtttaccg gagtagtccc gatattggtt gaactcgacg gcgatgtcaa
     5761 cggtcataaa ttcagtgtgt ccggcgaggg tgagggcgac gccacatacg gtaagctgac
     5821 gttgaagttc atatgcacca cgggcaagct gcccgtgcca tggccgacgt tggtcacgac
     5881 gctgacgtat ggtgtccagt gtttcagccg ttaccccgat catatgaagc agcatgactt
     5941 tttcaagtcg gcgatgccgg agggatacgt tcaagagagg accattttct tcaaggatga
     6001 cggcaactat aagacgcgag cggaggtgaa atttgaaggc gacacactcg ttaaccgtat
     6061 tgagttgaag ggcattgatt ttaaggagga tggcaacatt ctgggccata agttggaata
     6121 caactacaac tcgcataatg tgtatataat ggcagataag cagaaaaacg gaataaaggt
     6181 taacttcaag attcgccaca acatagagga cggttccgtg caacttgcag atcattacca
     6241 acagaacaca cccattggag atggcccagt tctcttgcca gacaatcact acctttccac
     6301 acagtccgcg ttgagcaagg accccaatga aaagcgggac cacatggtgt tgctggagtt
     6361 tgtgaccgca gctggtatta cacacggcat ggatgagctc tacaaggagg gccgcggcag
     6421 cctgctgacc tgcggcgatg tggaggagaa ccccgggccc atggattaca acaaaaggag
     6481 tagtgtgagt acggtgccga atgctgctcc cattcgcgtg ggcttcgtgg gattgaacgc
     6541 ggctaagggt tgggccatta aaacgcatta tccagccata ctgcagctga gctcccagtt
     6601 ccaaataaca gcgctttatt cccccaagat cgagacgtcc attgcgacaa ttcagaggct
     6661 gaagttgtcc aacgccacag ctttcccaac actcgagagc ttcgcctcga gctccacgat
     6721 cgatatgatt gtgatcgcaa tccaggtggc ttcccactac gaggtggtaa tgcccctgct
     6781 cgagttttcg aagaacaatc ctaatctgaa gtatttgttt gtggaatggg cactggcctg
     6841 ctcgctcgat caggcggagt cgatctataa ggccgctgca gagcggggcg tgcaaacaat
     6901 aatcagtctg cagggacgaa agagcccgta cattttgcgg gcaaaagagc ttatctccca
     6961 gggctacatc ggcgatataa atagcatcga gattgcaggt aacggaggtt ggtacggtta
     7021 cgagcggccg gtgaaaagcc cgaaatacat ttacgagatc ggaaacggag ttgatctggt
     7081 gaccaccacg ttcggtcaca cgatagatat attgcagtac atgaccagtt cgtacttcag
     7141 ccgtatcaat gccatggtgt ttaacaacat tccagaacag gagctcattg atgagagggg
     7201 caatcgcctg ggccagcggg tcccaaagac tgtcccagat catctcttgt tccagggcac
     7261 gctgctcaac ggaaatgttc ctgtgtcctg ttcctttaag ggaggcaagc ctacgaaaaa
     7321 gttcaccaag aatctggtca tagatatcca tggtacaaag ggtgatctga agctggaggg
     7381 tgacgctgga tttgcggaaa tctcgaatct cgttctgtat tattcgggca cccgcgccaa
     7441 tgattttccc ctggccaatg gtcagcaggc acccttggac cccggctacg acgcaggcaa
     7501 ggagattatg gaggtgtatc acctccgaaa ctacaacgcg atcgttggta atatacaccg
     7561 gctctaccag agtatcagcg atttccactt caacaccaag aaaatccccg agcttccatc
     7621 gcagttcgta atgcaaggct tcgatttcga aggctttccc accctgatgg atgccttgat
     7681 cttgcatcgc ctgattgaaa gtgtctacaa gtcgaatatg atgggcagta ccctgaatgt
     7741 ttccaacatc tcccactact cgctttaata ataaaatgaa tcgtagatac tgaaaaaccc
     7801 cgcaagttca cttcaactgt gcatcgtgca ccatctcaat ttctttcatt tatacatcgt
     7861 tttgccttct tttatgtaac tatactcctc taagtttcaa tcttggccat gtaacctctg
     7921 atctatagaa ttttttaaat gactagaatt aatgcccatc ttttttttgg acctaaattc
     7981 ttcatgaaaa tatattacga gggcttattc agaagcttat cgataccgtc gactaaagcc
     8041 aaatagaaat tattcagttc tggcttaagt ttttaaaagt gatattattt atttggttgt
     8101 aaccaaccaa aagaatgtaa ataactaata cataattatg ttagttttaa gttagcaaca
     8161 aattgatttt agctatatta gctacttggt taataaatag aatatattta tttaaagata
     8221 attcgttttt attgtcaggg agtgagtttg cttaaaaact cgtttggatc cggaggtagc
     8281 ggtggaagcg gaggttccgg caggtaagtt attgaacaat ggcatcaaat gccttcatca
     8341 tcactaccct ttagccctta agaccccaca atgaccttac ccactcagag aaaaaagtaa
     8401 atatgaaagc ccatttgaac ttctaagctt gtcgaccatc atgatggacc agatgggtga
     8461 ggtggagtac gcgcccgggg agcccaaggg cacgccctgg cacccgcacc gcggcttcga
     8521 gaccgtgacc tacatcatgc atggtaccca gcttttgttc cctttagtga gggttaattg
     8581 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     8641 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     8701 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcgg
//
