LOCUS       sequence-104090-        8748 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
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(1)-T2A-3X Gal80-Hsp70
FEATURES             Location/Qualifiers
     source          1..8748
                     /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             4278..4304
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4308..4331
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4332..4388
                     /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             5694..6407
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             6408..6461
                     /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 tccgcaggtg
     2881 gcggaggttc cggtggaagc ggaggtagcg gcggatccga gggccgcggc agcctgctga
     2941 cctgcggcga tgtggaggag aaccccgggc ccatggatta caacaaaagg agtagtgtga
     3001 gtacggtgcc gaatgctgct cccattcgcg tgggcttcgt gggattgaac gcggctaagg
     3061 gttgggccat taaaacgcat tatccagcca tactgcagct gagctcccag ttccaaataa
     3121 cagcgcttta ttcccccaag atcgagacgt ccattgcgac aattcagagg ctgaagttgt
     3181 ccaacgccac agctttccca acactcgaga gcttcgcctc gagctccacg atcgatatga
     3241 ttgtgatcgc aatccaggtg gcttcccact acgaggtggt aatgcccctg ctcgagtttt
     3301 cgaagaacaa tcctaatctg aagtatttgt ttgtggaatg ggcactggcc tgctcgctcg
     3361 atcaggcgga gtcgatctat aaggccgctg cagagcgggg cgtgcaaaca ataatcagtc
     3421 tgcagggacg aaagagcccg tacattttgc gggcaaaaga gcttatctcc cagggctaca
     3481 tcggcgatat aaatagcatc gagattgcag gtaacggagg ttggtacggt tacgagcggc
     3541 cggtgaaaag cccgaaatac atttacgaga tcggaaacgg agttgatctg gtgaccacca
     3601 cgttcggtca cacgatagat atattgcagt acatgaccag ttcgtacttc agccgtatca
     3661 atgccatggt gtttaacaac attccagaac aggagctcat tgatgagagg ggcaatcgcc
     3721 tgggccagcg ggtcccaaag actgtcccag atcatctctt gttccagggc acgctgctca
     3781 acggaaatgt tcctgtgtcc tgttccttta agggaggcaa gcctacgaaa aagttcacca
     3841 agaatctggt catagatatc catggtacaa agggtgatct gaagctggag ggtgacgctg
     3901 gatttgcgga aatctcgaat ctcgttctgt attattcggg cacccgcgcc aatgattttc
     3961 ccctggccaa tggtcagcag gcacccttgg accccggcta cgacgcaggc aaggagatta
     4021 tggaggtgta tcacctccga aactacaacg cgatcgttgg taatatacac cggctctacc
     4081 agagtatcag cgatttccac ttcaacacca agaaaatccc cgagcttcca tcgcagttcg
     4141 taatgcaagg cttcgatttc gaaggctttc ccaccctgat ggatgccttg atcttgcatc
     4201 gcctgattga aagtgtctac aagtcgaata tgatgggcag taccctgaat gtttccaaca
     4261 tctcccacta ctcgctttac ccctacgatg tgcccgatta cgccggcgat tacaaggacg
     4321 atgacgacaa ggccaccaac ttcagcctgc tgaagcaggc cggcgatgtg gaggagaacc
     4381 ccgggcccat ggattacaac aaaaggagta gtgtgagtac ggtgccgaat gctgctccca
     4441 ttcgcgtggg cttcgtggga ttgaacgcgg ctaagggttg ggccattaaa acgcattatc
     4501 cagccatact gcagctgagc tcccagttcc aaataacagc gctttattcc cccaagatcg
     4561 agacgtccat tgcgacaatt cagaggctga agttgtccaa cgccacagct ttcccaacac
     4621 tcgagagctt cgcctcgagc tccacgatcg atatgattgt gatcgcaatc caggtggctt
     4681 cccactacga ggtggtaatg cccctgctcg agttttcgaa gaacaatcct aatctgaagt
     4741 atttgtttgt ggaatgggca ctggcctgct cgctcgatca ggcggagtcg atctataagg
     4801 ccgctgcaga gcggggcgtg caaacaataa tcagtctgca gggacgaaag agcccgtaca
     4861 ttttgcgggc aaaagagctt atctcccagg gctacatcgg cgatataaat agcatcgaga
     4921 ttgcaggtaa cggaggttgg tacggttacg agcggccggt gaaaagcccg aaatacattt
     4981 acgagatcgg aaacggagtt gatctggtga ccaccacgtt cggtcacacg atagatatat
     5041 tgcagtacat gaccagttcg tacttcagcc gtatcaatgc catggtgttt aacaacattc
     5101 cagaacagga gctcattgat gagaggggca atcgcctggg ccagcgggtc ccaaagactg
     5161 tcccagatca tctcttgttc cagggcacgc tgctcaacgg aaatgttcct gtgtcctgtt
     5221 cctttaaggg aggcaagcct acgaaaaagt tcaccaagaa tctggtcata gatatccatg
     5281 gtacaaaggg tgatctgaag ctggagggtg acgctggatt tgcggaaatc tcgaatctcg
     5341 ttctgtatta ttcgggcacc cgcgccaatg attttcccct ggccaatggt cagcaggcac
     5401 ccttggaccc cggctacgac gcaggcaagg agattatgga ggtgtatcac ctccgaaact
     5461 acaacgcgat cgttggtaat atacaccggc tctaccagag tatcagcgat ttccacttca
     5521 acaccaagaa aatccccgag cttccatcgc agttcgtaat gcaaggcttc gatttcgaag
     5581 gctttcccac cctgatggat gccttgatct tgcatcgcct gattgaaagt gtctacaagt
     5641 cgaatatgat gggcagtacc ctgaatgttt ccaacatctc ccactactcg cttatgtcca
     5701 aaggtgaaga actgtttacc ggagtagtcc cgatattggt tgaactcgac ggcgatgtca
     5761 acggtcataa attcagtgtg tccggcgagg gtgagggcga cgccacatac ggtaagctga
     5821 cgttgaagtt catatgcacc acgggcaagc tgcccgtgcc atggccgacg ttggtcacga
     5881 cgctgacgta tggtgtccag tgtttcagcc gttaccccga tcatatgaag cagcatgact
     5941 ttttcaagtc ggcgatgccg gagggatacg ttcaagagag gaccattttc ttcaaggatg
     6001 acggcaacta taagacgcga gcggaggtga aatttgaagg cgacacactc gttaaccgta
     6061 ttgagttgaa gggcattgat tttaaggagg atggcaacat tctgggccat aagttggaat
     6121 acaactacaa ctcgcataat gtgtatataa tggcagataa gcagaaaaac ggaataaagg
     6181 ttaacttcaa gattcgccac aacatagagg acggttccgt gcaacttgca gatcattacc
     6241 aacagaacac acccattgga gatggcccag ttctcttgcc agacaatcac tacctttcca
     6301 cacagtccgc gttgagcaag gaccccaatg aaaagcggga ccacatggtg ttgctggagt
     6361 ttgtgaccgc agctggtatt acacacggca tggatgagct ctacaaggag ggccgcggca
     6421 gcctgctgac ctgcggcgat gtggaggaga accccgggcc catggattac aacaaaagga
     6481 gtagtgtgag tacggtgccg aatgctgctc ccattcgcgt gggcttcgtg ggattgaacg
     6541 cggctaaggg ttgggccatt aaaacgcatt atccagccat actgcagctg agctcccagt
     6601 tccaaataac agcgctttat tcccccaaga tcgagacgtc cattgcgaca attcagaggc
     6661 tgaagttgtc caacgccaca gctttcccaa cactcgagag cttcgcctcg agctccacga
     6721 tcgatatgat tgtgatcgca atccaggtgg cttcccacta cgaggtggta atgcccctgc
     6781 tcgagttttc gaagaacaat cctaatctga agtatttgtt tgtggaatgg gcactggcct
     6841 gctcgctcga tcaggcggag tcgatctata aggccgctgc agagcggggc gtgcaaacaa
     6901 taatcagtct gcagggacga aagagcccgt acattttgcg ggcaaaagag cttatctccc
     6961 agggctacat cggcgatata aatagcatcg agattgcagg taacggaggt tggtacggtt
     7021 acgagcggcc ggtgaaaagc ccgaaataca tttacgagat cggaaacgga gttgatctgg
     7081 tgaccaccac gttcggtcac acgatagata tattgcagta catgaccagt tcgtacttca
     7141 gccgtatcaa tgccatggtg tttaacaaca ttccagaaca ggagctcatt gatgagaggg
     7201 gcaatcgcct gggccagcgg gtcccaaaga ctgtcccaga tcatctcttg ttccagggca
     7261 cgctgctcaa cggaaatgtt cctgtgtcct gttcctttaa gggaggcaag cctacgaaaa
     7321 agttcaccaa gaatctggtc atagatatcc atggtacaaa gggtgatctg aagctggagg
     7381 gtgacgctgg atttgcggaa atctcgaatc tcgttctgta ttattcgggc acccgcgcca
     7441 atgattttcc cctggccaat ggtcagcagg cacccttgga ccccggctac gacgcaggca
     7501 aggagattat ggaggtgtat cacctccgaa actacaacgc gatcgttggt aatatacacc
     7561 ggctctacca gagtatcagc gatttccact tcaacaccaa gaaaatcccc gagcttccat
     7621 cgcagttcgt aatgcaaggc ttcgatttcg aaggctttcc caccctgatg gatgccttga
     7681 tcttgcatcg cctgattgaa agtgtctaca agtcgaatat gatgggcagt accctgaatg
     7741 tttccaacat ctcccactac tcgctttaat aataaaatga atcgtagata ctgaaaaacc
     7801 ccgcaagttc acttcaactg tgcatcgtgc accatctcaa tttctttcat ttatacatcg
     7861 ttttgccttc ttttatgtaa ctatactcct ctaagtttca atcttggcca tgtaacctct
     7921 gatctataga attttttaaa tgactagaat taatgcccat cttttttttg gacctaaatt
     7981 cttcatgaaa atatattacg agggcttatt cagaagctta tcgataccgt cgactaaagc
     8041 caaatagaaa ttattcagtt ctggcttaag tttttaaaag tgatattatt tatttggttg
     8101 taaccaacca aaagaatgta aataactaat acataattat gttagtttta agttagcaac
     8161 aaattgattt tagctatatt agctacttgg ttaataaata gaatatattt atttaaagat
     8221 aattcgtttt tattgtcagg gagtgagttt gcttaaaaac tcgtttggat ccggaggtag
     8281 cggtggaagc ggaggttccg gaggtaagtt 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
//
