LOCUS       sequence-357882-        7215 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 7215)
  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 7215)
  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{3P3-eGFP::GW,attB}
FEATURES             Location/Qualifiers
     source          1..7215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    47..80
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    121..190
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(409..429)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(409..427)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(448..464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(448..464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(461..483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    472..488
                     /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(496..502,503..520,521..526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    541..562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(679..696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(850..1438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(930..949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1609..2400,2401..2469))
                     /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     2232..2251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2470..2574)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2601..3056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2688..2707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2898..2919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3182..3204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3196..3213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3197..3213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3223..3242
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3223..3241
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3550..3559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3556..3558,3559..3561,3562..4074)
                     /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(3601..3622)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3862..3881)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    4309..4433
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4474..4779)
                     /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(4576..4595)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5121..5780)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5695..5714
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5834..5840,5841..5858,5859..5864))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5889..6013)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ccataaatct gaaacaatta ggcaaaacca tgcgaagctc tctagtataa cttcgtataa
       61 tgtatgctat acgaagttct cgaggggggg cccggtaccg tcgacgatgt aggtcacggt
      121 ctcgaagccg cggtgcgggt gccagggcgt gcccttgggc tccccgggcg cgtactccac
      181 ctcacccatc tggtccatca tgatgaacgg gtcgaggtgg cggtagttga tcccggcgaa
      241 cgcgcggcgc gccgggaagc cctcgccctc gaaaccgctg ggcgcggtgg tcacggtgag
      301 cacgggacgt gcgacggcgt cggcgggtgc ggatacgcgg ggcagcgtca gcgggttctc
      361 gacggtcacg gcgggcatgt cgacactagt ggtacccagc ttttgttccc tttagtgagg
      421 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      481 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      541 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      601 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      661 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      721 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      781 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      841 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      901 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      961 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1021 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1081 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1141 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1201 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1261 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1321 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1381 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1441 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1501 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1561 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1621 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1681 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1741 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1801 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1861 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1921 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1981 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2041 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2101 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2161 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2221 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2281 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2341 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2401 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2461 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2521 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2581 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2941 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3001 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat
     3061 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3121 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3181 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     3241 ggcgaattgg agctccaccg cggtggccat taatcatatg caatggttaa ttcgagctcg
     3301 cccggggatc taattcaatt agagactaat tcaattagag ctaattcaat taggatccaa
     3361 gcttatcgat ttcgattaac cctcgaccgc cggagtataa atagaggcgc ttcgtctacg
     3421 gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa
     3481 taaacaagcg cagctgaaca agctaaacaa tcggggtacc gctagagtcg acggtacgat
     3541 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaggtgag tagtacatgt
     4201 gcatacatct taagttcact tgatctatag gaactgcgat tgcaacatca aattgtctgc
     4261 ggcgtgagaa ctgcgaccca caaaaatccc aaaccgcggc cgtttatcac cactttgtac
     4321 aagaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     4381 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggtcgacc
     4441 tgcagactgg ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat
     4501 ggcgtttttg atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga
     4561 gaccggcaca ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac
     4621 caccgggtaa agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat
     4681 caccatccgt cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata
     4741 acggctctct cttttatagg tgtaaacctt aaactgcatt tcaccagccc ctgttctcgt
     4801 cagcaaaaga gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt
     4861 ccgctttcca gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca
     4921 gaccggagat attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt
     4981 cactgtaata cgctgcttca tagcatacct ctttttgaca tacttcgggt atacatatca
     5041 gtatatattc ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt
     5101 agtaagccgg atccacgcgt ttacgccccg ccctgccact catcgcagta ctgttgtaat
     5161 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     5221 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     5281 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     5341 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     5401 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     5461 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     5521 ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc
     5581 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     5641 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     5701 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     5761 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga cggatcctaa
     5821 ctcaaaatcc acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5881 gcggccgcca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     5941 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagcttttt
     6001 tgtacaaact tgtgataaac gaattcttga agacgaaagg gcacaaacaa atagtgacac
     6061 gaaacagatt attctggtag ctgttctcgc tatataagac aatttttgag atcatatcat
     6121 gatcaagaca tctaaaggca ttcattttcg actatattct tttttacaaa aaatataaca
     6181 accagatatt ttaagctgat cctagatgca caaaaaataa ataaaagtat aaacctactt
     6241 cgtaggatac ttcggggtac tttttgttcg gggttagatg agcataacgc ttgtagttga
     6301 tatttgagat cccctatcat tgcaggaccc aaacgagaag cgtgatcaca tggtcctcct
     6361 cgagttcgtg accgccgccg gcatcaccct gggcatggac gagctgtaca agtaagtata
     6421 gtgccttgac cgagctcgcc cactaagcgt cgcgccactt caacgctcga tgggagcgtc
     6481 attggtgggc ggggtaaccg tcgaaatcag tgtttacgct tccaatcgca acaaaaaatt
     6541 cactgcaaca ctgaaaagca tacgaaaacg atgaagattg tacgagaaac cataaagtat
     6601 tttatccaca aagacacgta tagcagaaaa gccaagttaa ctcggcgata agttgtgtac
     6661 acaagaataa aatcggccag attcagtgtt gtcagaaata agaaaacccc actatgtttt
     6721 tctttgcctt ttctttctcc cagcgatcat tcatttcgtg gtgaaagaac ggggtcattg
     6781 cacggagttt cgactgcggg aaagcagagc tgccgttcac ttcgtctata attagcgctt
     6841 tctattttcc ccgattcggg ccgctgctgc gcttttccgc ctgctgtttg tggcaagtgt
     6901 agcagcaggc tgtgcacgca gtgtggcatg cacttggctt tccaccgttg gtatcgattc
     6961 tctgggacga tgagtcattc ctttcggggc cacagcataa tcgttgccag ctcaccgaaa
     7021 tggtgacttc atttcttaac tgccgtcaag catgcgattg tacatacata catatttata
     7081 tatgtacata tttatgtgac tatggtaggt cgatataata gcaatcaacg caagcaaatg
     7141 tgtcagtcct gcttacagga acgattctat ttagtaattt tcgttgtata aagtaattat
     7201 gtatgtatgt aagcc
//
