LOCUS       sequence-357799-        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,attB}
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    213..246
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    287..356
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(575..595)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(575..593)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(614..630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(614..630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(627..649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    638..654
                     /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(662..668,669..686,687..692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    707..728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(845..862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1016..1604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1096..1115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1775..2566,2567..2635))
                     /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     2398..2417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2636..2740)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2767..3222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2854..2873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3064..3085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3348..3370
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3362..3379
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3363..3379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3389..3408
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3389..3407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3458..3940
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        3944..4535
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     regulatory      4716..4725
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4722..4724,4725..4727,4728..5240)
                     /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(4767..4788)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5028..5047)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    5475..5499
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    5516..5549
                     /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(5570..5594)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gcatgcgatt gtacatacat acatatttat atatgtacat atttatgtga ctatggtagg
       61 tcgatataat agcaatcaac gcaagcaaat gtgtcagtcc tgcttacagg aacgattcta
      121 tttagtaatt ttcgttgtat aaagtaatta tgtatgtatg taagccccat aaatctgaaa
      181 caattaggca aaaccatgcg aagctctcta gtataacttc gtataatgta tgctatacga
      241 agttctcgag ggggggcccg gtaccgtcga cgatgtaggt cacggtctcg aagccgcggt
      301 gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta ctccacctca cccatctggt
      361 ccatcatgat gaacgggtcg aggtggcggt agttgatccc ggcgaacgcg cggcgcgccg
      421 ggaagccctc gccctcgaaa ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga
      481 cggcgtcggc gggtgcggat acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg
      541 gcatgtcgac actagtggta cccagctttt gttcccttta gtgagggtta attgcgcgct
      601 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      661 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      721 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      781 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      841 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      901 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      961 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1021 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1081 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1141 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1201 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1261 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1321 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1381 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1441 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1501 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1561 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1621 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1681 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1741 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1801 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1861 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1921 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1981 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2041 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2101 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2161 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2221 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2281 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2341 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2401 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2461 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2521 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2581 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2641 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2701 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2761 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2941 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3061 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3121 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3181 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
     3241 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3301 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3361 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct
     3421 ccaccgcggt ggccattaat ccgagctctt acgcgcccgc gtaaaacaca atcaagtatg
     3481 agtcataagc tgatgtcatg ttttgcacac ggctcataac cgaactggct ttacgagtag
     3541 aattctactt gtaacgcacg atcagtggat gatgtcattt gtttttcaaa tcgagatgat
     3601 gtcatgtttt gcacacggct cataaactcg ctttacgagt agaattctac gtgtaacgca
     3661 cgatcgattg atgagtcatt tgttttgcaa tatgatatca tacaatatga ctcatttgtt
     3721 tttcaaaacc gaacttgatt tacgggtaga attctacttg taaagcacaa tcaaaaagat
     3781 gatgtcattt gtttttcaaa actgaactcg ctttacgagt agaattctac gtgtaaaaca
     3841 caatcaagaa atgatgtcat ttgttataaa aataaaagct gatgtcatgt tttgcacatg
     3901 gctcataact aaactcgctt tacgggtaga attctacgcg cgtcgatgtc tttgtgatgc
     3961 gcgcgacatt tttgtaggtt attgataaaa tgaacggata cgttgcccga cattatcatt
     4021 aaatccttgg cgtagaattt gtcgggtcca ttgtccgtgt gcgctagcat gcccgtaacg
     4081 gacctcgtac ttttggcttc aaaggttttg cgcacagaca aaatgtgcca cacttgcagc
     4141 tctgcatgtg tgcgcgttac cacaaatccc aacggcgcag tgtacttgtt gtatgcaaat
     4201 aaatctcgat aaaggcgcgg cgcgcgaatg cagctgatca cgtacgctcc tcgtgttccg
     4261 ttcaaggacg gtgttatcga cctcagatta atgtttatcg gccgactgtt ttcgtatccg
     4321 ctcaccaaac gcgtttttgc attaacattg tatgtcggcg gatgttctat atctaatttg
     4381 aataaataaa cgataaccgc gttggtttta gagggcataa taaaagaaat attgttatcg
     4441 tgttcgccat tagggcagta taaattgacg ttcatgttgg atattgtttc agttgcaagt
     4501 tgacactggc ggcgacaaga tcgtgaacaa ccaagtgacc attaaccctc gaccgccgga
     4561 gtataaatag aggcgcttcg tctacggagc gacaattcaa ttcaaacaag caaagtgaac
     4621 acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc tgaacaagct aaacaatcgg
     4681 ggtaccgcta gagtcgacgg tacgatccac cggtcgccac catggtgagc aagggcgagg
     4741 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     4801 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     4861 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     4921 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4981 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     5041 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     5101 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     5161 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     5221 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     5281 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     5341 ccctgagcaa ggtgagtagt acatgtgcat acatcttaag ttcacttgat ctataggaac
     5401 tgcgattgca acatcaaatt gtctgcggcg tgagaactgc gacccacaaa aatcccaaac
     5461 cgcggccgtt tatcaccact ttgtacaaga aagctgggtc ggcgcgccca cccttgaagt
     5521 tcctattctc tagaaagtat aggaacttcg gtgaaggggg cggccgcgga gcctgctttt
     5581 ttgtacaaac ttgtgataaa cgaattcttg aagacgaaag ggcacaaaca aatagtgaca
     5641 cgaaacagat tattctggta gctgttctcg ctatataaga caatttttga gatcatatca
     5701 tgatcaagac atctaaaggc attcattttc gactatattc ttttttacaa aaaatataac
     5761 aaccagatat tttaagctga tcctagatgc acaaaaaata aataaaagta taaacctact
     5821 tcgtaggata cttcggggta ctttttgttc ggggttagat gagcataacg cttgtagttg
     5881 atatttgaga tcccctatca ttgcaggacc caaacgagaa gcgtgatcac atggtcctcc
     5941 tcgagttcgt gaccgccgcc ggcatcaccc tgggcatgga cgagctgtac aagtaagtat
     6001 agtgccttga ccgagctcgc ccactaagcg tcgcgccact tcaacgctcg atgggagcgt
     6061 cattggtggg cggggtaacc gtcgaaatca gtgtttacgc ttccaatcgc aacaaaaaat
     6121 tcactgcaac actgaaaagc atacgaaaac gatgaagatt gtacgagaaa ccataaagta
     6181 ttttatccac aaagacacgt atagcagaaa agccaagtta actcggcgat aagttgtgta
     6241 cacaagaata aaatcggcca gattcagtgt tgtcagaaat aagaaaaccc cactatgttt
     6301 ttctttgcct tttctttctc ccagcgatca ttcatttcgt ggtgaaagaa cggggtcatt
     6361 gcacggagtt tcgactgcgg gaaagcagag ctgccgttca cttcgtctat aattagcgct
     6421 ttctattttc cccgattcgg gccgctgctg cgcttttccg cctgctgttt gtggcaagtg
     6481 tagcagcagg ctgtgcacgc agtgtggcat gcacttggct ttccaccgtt ggtatcgatt
     6541 ctctgggacg atgagtcatt cctttcgggg ccacagcata atcgttgcca gctcaccgaa
     6601 atggtgactt catttcttaa ctgccgtcaa
//
