LOCUS       sequence-357827-        6206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent reporter gene with yeast recombination site within 
            intron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6206)
  AUTHORS   Stern DL
  TITLE     Transgenic tools for targeted chromosome rearrangements allow 
            construction of balancer chromosomes in non-melanogaster Drosophila 
            species.
  JOURNAL   G3 (Bethesda). 2022 Feb 10. pii: 6526388. doi: 
            10.1093/g3journal/jkac030.
  PUBMED    35143616
REFERENCE   2  (bases 1 to 6206)
  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"; journalName: "G3 
            (Bethesda). 2022 Feb 10. pii: 6526388. doi: 
            10.1093/g3journal/jkac030."
COMMENT     Sequence Label: p{Mhc-F4-678-eGFP::FRT,attB}
FEATURES             Location/Qualifiers
     source          1..6206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      269..278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(275..277,278..280,281..793)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(320..341)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(581..600)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    1028..1052
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    1069..1102
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(1123..1147)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    2396..2429
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2470..2539
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2758..2778)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2758..2776)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2797..2813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2797..2813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2810..2832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2821..2837
                     /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(2845..2851,2852..2869,2870..2875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2890..2911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3028..3045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3199..3787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3279..3298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3958..4749,4750..4818))
                     /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     4581..4600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4819..4923)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4950..5405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5037..5056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5247..5268
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5531..5553
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5545..5562
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5546..5562
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5572..5591
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5572..5590
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccgaaaatcc gtatatatgc aaagcattct gatatatata tataagtggt atacatatac
       61 atattcgtat cggatccgtg atctatattt aggattaacc ctcgaccgcc ggagtataaa
      121 tagaggcgct tcgtctacgg agcgacaatt caattcaaac aagcaaagtg aacacgtcgc
      181 taagcgaaag ctaagcaaat aaacaagcgc agctgaacaa gctaaacaat cggggtaccg
      241 ctagagtcga cggtacgatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
      301 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      361 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      421 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      481 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      541 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      601 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      661 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      721 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
      781 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
      841 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
      901 caaggtgagt agtacatgtg catacatctt aagttcactt gatctatagg aactgcgatt
      961 gcaacatcaa attgtctgcg gcgtgagaac tgcgacccac aaaaatccca aaccgcggcc
     1021 gtttatcacc actttgtaca agaaagctgg gtcggcgcgc ccacccttga agttcctatt
     1081 ctctagaaag tataggaact tcggtgaagg gggcggccgc ggagcctgct tttttgtaca
     1141 aacttgtgat aaacgaattc ttgaagacga aagggcacaa acaaatagtg acacgaaaca
     1201 gattattctg gtagctgttc tcgctatata agacaatttt tgagatcata tcatgatcaa
     1261 gacatctaaa ggcattcatt ttcgactata ttctttttta caaaaaatat aacaaccaga
     1321 tattttaagc tgatcctaga tgcacaaaaa ataaataaaa gtataaacct acttcgtagg
     1381 atacttcggg gtactttttg ttcggggtta gatgagcata acgcttgtag ttgatatttg
     1441 agatccccta tcattgcagg acccaaacga gaagcgtgat cacatggtcc tcctcgagtt
     1501 cgtgaccgcc gccggcatca ccctgggcat ggacgagctg tacaagtaag tatagtgcct
     1561 tgaccgagct cgcccactaa gcgtcgcgcc acttcaacgc tcgatgggag cgtcattggt
     1621 gggcggggta accgtcgaaa tcagtgttta cgcttccaat cgcaacaaaa aattcactgc
     1681 aacactgaaa agcatacgaa aacgatgaag attgtacgag aaaccataaa gtattttatc
     1741 cacaaagaca cgtatagcag aaaagccaag ttaactcggc gataagttgt gtacacaaga
     1801 ataaaatcgg ccagattcag tgttgtcaga aataagaaaa ccccactatg tttttctttg
     1861 ccttttcttt ctcccagcga tcattcattt cgtggtgaaa gaacggggtc attgcacgga
     1921 gtttcgactg cgggaaagca gagctgccgt tcacttcgtc tataattagc gctttctatt
     1981 ttccccgatt cgggccgctg ctgcgctttt ccgcctgctg tttgtggcaa gtgtagcagc
     2041 aggctgtgca cgcagtgtgg catgcacttg gctttccacc gttggtatcg attctctggg
     2101 acgatgagtc attcctttcg gggccacagc ataatcgttg ccagctcacc gaaatggtga
     2161 cttcatttct taactgccgt caagcatgcg attgtacata catacatatt tatatatgta
     2221 catatttatg tgactatggt aggtcgatat aatagcaatc aacgcaagca aatgtgtcag
     2281 tcctgcttac aggaacgatt ctatttagta attttcgttg tataaagtaa ttatgtatgt
     2341 atgtaagccc cataaatctg aaacaattag gcaaaaccat gcgaagctct ctagtataac
     2401 ttcgtataat gtatgctata cgaagttctc gagggggggc ccggtaccgt cgacgatgta
     2461 ggtcacggtc tcgaagccgc ggtgcgggtg ccagggcgtg cccttgggct ccccgggcgc
     2521 gtactccacc tcacccatct ggtccatcat gatgaacggg tcgaggtggc ggtagttgat
     2581 cccggcgaac gcgcggcgcg ccgggaagcc ctcgccctcg aaaccgctgg gcgcggtggt
     2641 cacggtgagc acgggacgtg cgacggcgtc ggcgggtgcg gatacgcggg gcagcgtcag
     2701 cgggttctcg acggtcacgg cgggcatgtc gacactagtg gtacccagct tttgttccct
     2761 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2821 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2881 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2941 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3001 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4921 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     4981 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5041 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5101 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5161 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5221 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5281 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5341 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5401 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5461 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     5521 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
     5581 tcactatagg gcgaattgga gctccaccgc ggtggccatt aatatactcg aattcgttgt
     5641 cggctcatat acatgggcga aataatttcg aatatgtttt aaaaataacc aaagacatta
     5701 gaaagagatc gccaatactt atacattatg tctatgtgtg ccatgtggta gcatgagcca
     5761 aaaagcttct cgaaattacg aattacatat agacgaatat gtatgtgact ttagtttcga
     5821 aataatttcg agaattttaa aaataacgca ttcgttaaaa gttcgcgtca attcgaatcg
     5881 aattttcgat tgtcgattta gtgtggattg tcgaaaatcg ttccgccttc gaagtttact
     5941 gaaaggaatc attgcgatct cgtgaattgc ttgtatgaga acacgccacc atatcgagat
     6001 acctgttcaa aatatcgaaa tcgcgttcca aacggtagta gttaccagtt gagtgagttg
     6061 tggtgacagt ttaacctctc tacatttgta ataataagca ttgttttggc gtatcaatag
     6121 tctgtagatt ttttcacaaa gaaacctgga gaaatgtaca caaatatcta aatgaaaatt
     6181 ggcgttacgt aacaaatacc aacgcc
//
