LOCUS       sequence-357763-        6235 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 6235)
  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 6235)
  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::KD,attB}
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      330..339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(336..338,339..341,342..854)
                     /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(381..402)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(642..661)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    1089..1113
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1211..1235)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    2484..2517
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2558..2627
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2846..2866)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2846..2864)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2885..2901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2885..2901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2898..2920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2909..2925
                     /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(2933..2939,2940..2957,2958..2963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2978..2999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3116..3133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3287..3875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3367..3386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4046..4837,4838..4906))
                     /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     4669..4688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4907..5011)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5038..5493
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5125..5144)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5335..5356
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5619..5641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5633..5650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5634..5650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5660..5679
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5660..5678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggagaaatgt acacaaatat ctaaatgaaa attggcgtta cgtaacaaat accaacgccc
       61 cgaaaatccg tatatatgca aagcattctg atatatatat atataagtgg tatacatata
      121 catattcgta tcggatccgt gatctatatt taggattaac cctcgaccgc cggagtataa
      181 atagaggcgc ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg
      241 ctaagcgaaa gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tcggggtacc
      301 gctagagtcg acggtacgat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt
      361 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      421 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      481 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      541 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      601 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      661 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      721 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      781 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
      841 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
      901 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
      961 gcaaggtgag tagtacatgt gcatacatct taagttcact tgatctatag gaactgcgat
     1021 tgcaacatca aattgtctgc ggcgtgagaa ctgcgaccca caaaaatccc aaaccgcggc
     1081 cgtttatcac cactttgtac aagaaagctg ggtcggcgcg cccaccctta aacgatatca
     1141 gacatttgtc tgataatgaa gcattatcag acaaatgtct gatatcgttt ggtgaagggg
     1201 gcggccgcgg agcctgcttt tttgtacaaa cttgtgataa acgaattctt gaagacgaaa
     1261 gggcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc gctatataag
     1321 acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt cgactatatt
     1381 cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg cacaaaaaat
     1441 aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt cggggttaga
     1501 tgagcataac gcttgtagtt gatatttgag atcccctatc attgcaggac ccaaacgaga
     1561 agcgtgatca catggtcctc ctcgagttcg tgaccgccgc cggcatcacc ctgggcatgg
     1621 acgagctgta caagtaagta tagtgccttg accgagctcg cccactaagc gtcgcgccac
     1681 ttcaacgctc gatgggagcg tcattggtgg gcggggtaac cgtcgaaatc agtgtttacg
     1741 cttccaatcg caacaaaaaa ttcactgcaa cactgaaaag catacgaaaa cgatgaagat
     1801 tgtacgagaa accataaagt attttatcca caaagacacg tatagcagaa aagccaagtt
     1861 aactcggcga taagttgtgt acacaagaat aaaatcggcc agattcagtg ttgtcagaaa
     1921 taagaaaacc ccactatgtt tttctttgcc ttttctttct cccagcgatc attcatttcg
     1981 tggtgaaaga acggggtcat tgcacggagt ttcgactgcg ggaaagcaga gctgccgttc
     2041 acttcgtcta taattagcgc tttctatttt ccccgattcg ggccgctgct gcgcttttcc
     2101 gcctgctgtt tgtggcaagt gtagcagcag gctgtgcacg cagtgtggca tgcacttggc
     2161 tttccaccgt tggtatcgat tctctgggac gatgagtcat tcctttcggg gccacagcat
     2221 aatcgttgcc agctcaccga aatggtgact tcatttctta actgccgtca agcatgcgat
     2281 tgtacataca tacatattta tatatgtaca tatttatgtg actatggtag gtcgatataa
     2341 tagcaatcaa cgcaagcaaa tgtgtcagtc ctgcttacag gaacgattct atttagtaat
     2401 tttcgttgta taaagtaatt atgtatgtat gtaagcccca taaatctgaa acaattaggc
     2461 aaaaccatgc gaagctctct agtataactt cgtataatgt atgctatacg aagttctcga
     2521 gggggggccc ggtaccgtcg acgatgtagg tcacggtctc gaagccgcgg tgcgggtgcc
     2581 agggcgtgcc cttgggctcc ccgggcgcgt actccacctc acccatctgg tccatcatga
     2641 tgaacgggtc gaggtggcgg tagttgatcc cggcgaacgc gcggcgcgcc gggaagccct
     2701 cgccctcgaa accgctgggc gcggtggtca cggtgagcac gggacgtgcg acggcgtcgg
     2761 cgggtgcgga tacgcggggc agcgtcagcg ggttctcgac ggtcacggcg ggcatgtcga
     2821 cactagtggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     2881 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2941 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3001 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3061 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5041 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5101 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5161 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5221 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5281 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5341 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5401 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5461 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     5521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5641 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattggagc tccaccgcgg
     5701 tggccattaa tatactcgaa ttcgttgtcg gctcatatac atgggcgaaa taatttcgaa
     5761 tatgttttaa aaataaccaa agacattaga aagagatcgc caatacttat acattatgtc
     5821 tatgtgtgcc atgtggtagc atgagccaaa aagcttctcg aaattacgaa ttacatatag
     5881 acgaatatgt atgtgacttt agtttcgaaa taatttcgag aattttaaaa ataacgcatt
     5941 cgttaaaagt tcgcgtcaat tcgaatcgaa ttttcgattg tcgatttagt gtggattgtc
     6001 gaaaatcgtt ccgccttcga agtttactga aaggaatcat tgcgatctcg tgaattgctt
     6061 gtatgagaac acgccaccat atcgagatac ctgttcaaaa tatcgaaatc gcgttccaaa
     6121 cggtagtagt taccagttga gtgagttgtg gtgacagttt aacctctcta catttgtaat
     6181 aataagcatt gttttggcgt atcaatagtc tgtagatttt ttcacaaaga aacct
//
