LOCUS       sequence-357770-        6630 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 6630)
  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 6630)
  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{ie1-eGFP::FRT-RC,attB}
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    206..239
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    280..349
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(568..588)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(568..586)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(607..623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(620..642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    631..647
                     /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(655..661,662..679,680..685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    700..721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(838..855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1009..1597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1089..1108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1768..2559,2560..2628))
                     /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     2391..2410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2629..2733)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2760..3215
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2847..2866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3057..3078
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3341..3363
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3355..3372
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3356..3372
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3382..3401
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3382..3400
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3451..3933
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        3937..4528
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     regulatory      4709..4718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4715..4717,4718..4720,4721..5233)
                     /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(4760..4781)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5021..5040)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    5468..5492
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5509..5542)
                     /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(5563..5587)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 attgtacata catacatatt tatatatgta catatttatg tgactatggt aggtcgatat
       61 aatagcaatc aacgcaagca aatgtgtcag tcctgcttac aggaacgatt ctatttagta
      121 attttcgttg tataaagtaa ttatgtatgt atgtaagccc cataaatctg aaacaattag
      181 gcaaaaccat gcgaagctct ctagtataac ttcgtataat gtatgctata cgaagttctc
      241 gagggggggc ccggtaccgt cgacgatgta ggtcacggtc tcgaagccgc ggtgcgggtg
      301 ccagggcgtg cccttgggct ccccgggcgc gtactccacc tcacccatct ggtccatcat
      361 gatgaacggg tcgaggtggc ggtagttgat cccggcgaac gcgcggcgcg ccgggaagcc
      421 ctcgccctcg aaaccgctgg gcgcggtggt cacggtgagc acgggacgtg cgacggcgtc
      481 ggcgggtgcg gatacgcggg gcagcgtcag cgggttctcg acggtcacgg cgggcatgtc
      541 gacactagtg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
      601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
      661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
      721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
      841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
      901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     2761 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2821 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2881 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2941 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3001 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3061 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3121 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3181 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     3241 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     3301 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     3361 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc
     3421 ggtggccatt aatccgagct cttacgcgcc cgcgtaaaac acaatcaagt atgagtcata
     3481 agctgatgtc atgttttgca cacggctcat aaccgaactg gctttacgag tagaattcta
     3541 cttgtaacgc acgatcagtg gatgatgtca tttgtttttc aaatcgagat gatgtcatgt
     3601 tttgcacacg gctcataaac tcgctttacg agtagaattc tacgtgtaac gcacgatcga
     3661 ttgatgagtc atttgttttg caatatgata tcatacaata tgactcattt gtttttcaaa
     3721 accgaacttg atttacgggt agaattctac ttgtaaagca caatcaaaaa gatgatgtca
     3781 tttgtttttc aaaactgaac tcgctttacg agtagaattc tacgtgtaaa acacaatcaa
     3841 gaaatgatgt catttgttat aaaaataaaa gctgatgtca tgttttgcac atggctcata
     3901 actaaactcg ctttacgggt agaattctac gcgcgtcgat gtctttgtga tgcgcgcgac
     3961 atttttgtag gttattgata aaatgaacgg atacgttgcc cgacattatc attaaatcct
     4021 tggcgtagaa tttgtcgggt ccattgtccg tgtgcgctag catgcccgta acggacctcg
     4081 tacttttggc ttcaaaggtt ttgcgcacag acaaaatgtg ccacacttgc agctctgcat
     4141 gtgtgcgcgt taccacaaat cccaacggcg cagtgtactt gttgtatgca aataaatctc
     4201 gataaaggcg cggcgcgcga atgcagctga tcacgtacgc tcctcgtgtt ccgttcaagg
     4261 acggtgttat cgacctcaga ttaatgttta tcggccgact gttttcgtat ccgctcacca
     4321 aacgcgtttt tgcattaaca ttgtatgtcg gcggatgttc tatatctaat ttgaataaat
     4381 aaacgataac cgcgttggtt ttagagggca taataaaaga aatattgtta tcgtgttcgc
     4441 cattagggca gtataaattg acgttcatgt tggatattgt ttcagttgca agttgacact
     4501 ggcggcgaca agatcgtgaa caaccaagtg accattaacc ctcgaccgcc ggagtataaa
     4561 tagaggcgct tcgtctacgg agcgacaatt caattcaaac aagcaaagtg aacacgtcgc
     4621 taagcgaaag ctaagcaaat aaacaagcgc agctgaacaa gctaaacaat cggggtaccg
     4681 ctagagtcga cggtacgatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     4741 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4801 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4861 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4921 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4981 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     5041 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     5101 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     5161 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     5221 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     5281 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     5341 caaggtgagt agtacatgtg catacatctt aagttcactt gatctatagg aactgcgatt
     5401 gcaacatcaa attgtctgcg gcgtgagaac tgcgacccac aaaaatccca aaccgcggcc
     5461 gtttatcacc actttgtaca agaaagctgg gtcggcgcgc ccacccttga agttcctata
     5521 ctttctagag aataggaact tcggtgaagg gggcggccgc ggagcctgct tttttgtaca
     5581 aacttgtgat aaacgaattc ttgaagacga aagggcacaa acaaatagtg acacgaaaca
     5641 gattattctg gtagctgttc tcgctatata agacaatttt tgagatcata tcatgatcaa
     5701 gacatctaaa ggcattcatt ttcgactata ttctttttta caaaaaatat aacaaccaga
     5761 tattttaagc tgatcctaga tgcacaaaaa ataaataaaa gtataaacct acttcgtagg
     5821 atacttcggg gtactttttg ttcggggtta gatgagcata acgcttgtag ttgatatttg
     5881 agatccccta tcattgcagg acccaaacga gaagcgtgat cacatggtcc tcctcgagtt
     5941 cgtgaccgcc gccggcatca ccctgggcat ggacgagctg tacaagtaag tatagtgcct
     6001 tgaccgagct cgcccactaa gcgtcgcgcc acttcaacgc tcgatgggag cgtcattggt
     6061 gggcggggta accgtcgaaa tcagtgttta cgcttccaat cgcaacaaaa aattcactgc
     6121 aacactgaaa agcatacgaa aacgatgaag attgtacgag aaaccataaa gtattttatc
     6181 cacaaagaca cgtatagcag aaaagccaag ttaactcggc gataagttgt gtacacaaga
     6241 ataaaatcgg ccagattcag tgttgtcaga aataagaaaa ccccactatg tttttctttg
     6301 ccttttcttt ctcccagcga tcattcattt cgtggtgaaa gaacggggtc attgcacgga
     6361 gtttcgactg cgggaaagca gagctgccgt tcacttcgtc tataattagc gctttctatt
     6421 ttccccgatt cgggccgctg ctgcgctttt ccgcctgctg tttgtggcaa gtgtagcagc
     6481 aggctgtgca cgcagtgtgg catgcacttg gctttccacc gttggtatcg attctctggg
     6541 acgatgagtc attcctttcg gggccacagc ataatcgttg ccagctcacc gaaatggtga
     6601 cttcatttct taactgccgt caagcatgcg
//
