LOCUS       sequence-357884-        7791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent reporter gene with intron containing Gateway cloning 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  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 7791)
  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::GW,attB}
FEATURES             Location/Qualifiers
     source          1..7791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(379..381,382..384,385..897)
                     /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(424..445)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(685..704)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    1132..1256
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1297..1602)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1399..1418)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1944..2603)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2518..2537
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2657..2663,2664..2681,2682..2687))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2712..2836)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    4085..4118
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4159..4228
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(4447..4467)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4447..4465)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4486..4502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4486..4502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4499..4521)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4510..4526
                     /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(4534..4540,4541..4558,4559..4564))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4579..4600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4717..4734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4888..5476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4968..4987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5647..6438,6439..6507))
                     /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     6270..6289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6508..6612)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6639..7094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6726..6745)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6936..6957
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7220..7242
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7234..7251
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7235..7251
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7261..7280
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7261..7279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gttttggcgt atcaatagtc tgtagatttt ttcacaaaga aacctggaga aatgtacaca
       61 aatatctaaa tgaaaattgg cgttacgtaa caaataccaa cgccccgaaa atccgtatat
      121 atgcaaagca ttctgatata tatatataag tggtatacat atacatattc gtatcggatc
      181 cgtgatctat atttaggatt aaccctcgac cgccggagta taaatagagg cgcttcgtct
      241 acggagcgac aattcaattc aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc
      301 aaataaacaa gcgcagctga acaagctaaa caatcggggt accgctagag tcgacggtac
      361 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      421 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      481 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      541 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      601 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      661 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      721 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      781 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      841 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      901 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      961 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaggt gagtagtaca
     1021 tgtgcataca tcttaagttc acttgatcta taggaactgc gattgcaaca tcaaattgtc
     1081 tgcggcgtga gaactgcgac ccacaaaaat cccaaaccgc ggccgtttat caccactttg
     1141 tacaagaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
     1201 ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc actatggtcg
     1261 acctgcagac tggctgtgta taagggagcc tgacatttat attccccaga acatcaggtt
     1321 aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac
     1381 ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg
     1441 caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac tggccagggg
     1501 gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca caaacagacg
     1561 ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag cccctgttct
     1621 cgtcagcaaa agagccgttc atttcaataa accgggcgac ctcagccatc ccttcctgat
     1681 tttccgcttt ccagcgttcg gcacgcagac gacgggcttc attctgcatg gttgtgctta
     1741 ccagaccgga gatattgaca tcatatatgc cttgagcaac tgatagctgt cgctgtcaac
     1801 tgtcactgta atacgctgct tcatagcata cctctttttg acatacttcg ggtatacata
     1861 tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg ttatctggct
     1921 tttagtaagc cggatccacg cgtttacgcc ccgccctgcc actcatcgca gtactgttgt
     1981 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     2041 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     2101 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     2161 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     2221 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     2281 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     2341 acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca
     2401 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     2461 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     2521 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     2581 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgacggatcc
     2641 taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2701 aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
     2761 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt
     2821 ttttgtacaa acttgtgata aacgaattct tgaagacgaa agggcacaaa caaatagtga
     2881 cacgaaacag attattctgg tagctgttct cgctatataa gacaattttt gagatcatat
     2941 catgatcaag acatctaaag gcattcattt tcgactatat tcttttttac aaaaaatata
     3001 acaaccagat attttaagct gatcctagat gcacaaaaaa taaataaaag tataaaccta
     3061 cttcgtagga tacttcgggg tactttttgt tcggggttag atgagcataa cgcttgtagt
     3121 tgatatttga gatcccctat cattgcagga cccaaacgag aagcgtgatc acatggtcct
     3181 cctcgagttc gtgaccgccg ccggcatcac cctgggcatg gacgagctgt acaagtaagt
     3241 atagtgcctt gaccgagctc gcccactaag cgtcgcgcca cttcaacgct cgatgggagc
     3301 gtcattggtg ggcggggtaa ccgtcgaaat cagtgtttac gcttccaatc gcaacaaaaa
     3361 attcactgca acactgaaaa gcatacgaaa acgatgaaga ttgtacgaga aaccataaag
     3421 tattttatcc acaaagacac gtatagcaga aaagccaagt taactcggcg ataagttgtg
     3481 tacacaagaa taaaatcggc cagattcagt gttgtcagaa ataagaaaac cccactatgt
     3541 ttttctttgc cttttctttc tcccagcgat cattcatttc gtggtgaaag aacggggtca
     3601 ttgcacggag tttcgactgc gggaaagcag agctgccgtt cacttcgtct ataattagcg
     3661 ctttctattt tccccgattc gggccgctgc tgcgcttttc cgcctgctgt ttgtggcaag
     3721 tgtagcagca ggctgtgcac gcagtgtggc atgcacttgg ctttccaccg ttggtatcga
     3781 ttctctggga cgatgagtca ttcctttcgg ggccacagca taatcgttgc cagctcaccg
     3841 aaatggtgac ttcatttctt aactgccgtc aagcatgcga ttgtacatac atacatattt
     3901 atatatgtac atatttatgt gactatggta ggtcgatata atagcaatca acgcaagcaa
     3961 atgtgtcagt cctgcttaca ggaacgattc tatttagtaa ttttcgttgt ataaagtaat
     4021 tatgtatgta tgtaagcccc ataaatctga aacaattagg caaaaccatg cgaagctctc
     4081 tagtataact tcgtataatg tatgctatac gaagttctcg agggggggcc cggtaccgtc
     4141 gacgatgtag gtcacggtct cgaagccgcg gtgcgggtgc cagggcgtgc ccttgggctc
     4201 cccgggcgcg tactccacct cacccatctg gtccatcatg atgaacgggt cgaggtggcg
     4261 gtagttgatc ccggcgaacg cgcggcgcgc cgggaagccc tcgccctcga aaccgctggg
     4321 cgcggtggtc acggtgagca cgggacgtgc gacggcgtcg gcgggtgcgg atacgcgggg
     4381 cagcgtcagc gggttctcga cggtcacggc gggcatgtcg acactagtgg tacccagctt
     4441 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4561 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4621 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4681 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4741 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4801 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4861 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4921 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4981 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5041 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5101 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5161 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5221 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5281 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5341 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5401 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5581 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5641 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5821 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6061 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6301 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6481 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6541 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6601 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     6661 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     6721 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6781 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6841 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6901 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6961 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7021 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7081 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7141 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7201 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     7261 taatacgact cactataggg cgaattggag ctccaccgcg gtggccatta atatactcga
     7321 attcgttgtc ggctcatata catgggcgaa ataatttcga atatgtttta aaaataacca
     7381 aagacattag aaagagatcg ccaatactta tacattatgt ctatgtgtgc catgtggtag
     7441 catgagccaa aaagcttctc gaaattacga attacatata gacgaatatg tatgtgactt
     7501 tagtttcgaa ataatttcga gaattttaaa aataacgcat tcgttaaaag ttcgcgtcaa
     7561 ttcgaatcga attttcgatt gtcgatttag tgtggattgt cgaaaatcgt tccgccttcg
     7621 aagtttactg aaaggaatca ttgcgatctc gtgaattgct tgtatgagaa cacgccacca
     7681 tatcgagata cctgttcaaa atatcgaaat cgcgttccaa acggtagtag ttaccagttg
     7741 agtgagttgt ggtgacagtt taacctctct acatttgtaa taataagcat t
//
