LOCUS       sequence-357877-        8215 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 8215)
  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 8215)
  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::GW,attB}
FEATURES             Location/Qualifiers
     source          1..8215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    196..229
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    270..339
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(558..578)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(558..576)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(597..613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(597..613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(610..632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    621..637
                     /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(645..651,652..669,670..675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    690..711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(828..845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(999..1587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1079..1098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1758..2549,2550..2618))
                     /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     2381..2400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2619..2723)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2750..3205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2837..2856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3047..3068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3331..3353
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3345..3362
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3346..3362
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3372..3391
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3372..3390
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3441..3923
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        3927..4518
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     regulatory      4699..4708
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4705..4707,4708..4710,4711..5223)
                     /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(4750..4771)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5011..5030)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    5458..5582
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(5623..5928)
                     /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(5725..5744)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6270..6929)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6844..6863
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6983..6989,6990..7007,7008..7013))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(7038..7162)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 catacatatt tatatatgta catatttatg tgactatggt aggtcgatat aatagcaatc
       61 aacgcaagca aatgtgtcag tcctgcttac aggaacgatt ctatttagta attttcgttg
      121 tataaagtaa ttatgtatgt atgtaagccc cataaatctg aaacaattag gcaaaaccat
      181 gcgaagctct ctagtataac ttcgtataat gtatgctata cgaagttctc gagggggggc
      241 ccggtaccgt cgacgatgta ggtcacggtc tcgaagccgc ggtgcgggtg ccagggcgtg
      301 cccttgggct ccccgggcgc gtactccacc tcacccatct ggtccatcat gatgaacggg
      361 tcgaggtggc ggtagttgat cccggcgaac gcgcggcgcg ccgggaagcc ctcgccctcg
      421 aaaccgctgg gcgcggtggt cacggtgagc acgggacgtg cgacggcgtc ggcgggtgcg
      481 gatacgcggg gcagcgtcag cgggttctcg acggtcacgg cgggcatgtc gacactagtg
      541 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
      601 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      661 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      721 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      781 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
      841 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      901 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      961 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1021 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1081 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1141 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1201 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1261 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1321 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1381 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1441 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1501 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1561 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1621 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1681 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1741 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1801 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1861 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1921 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1981 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2041 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2101 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2161 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2221 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2281 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2341 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2401 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2461 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2521 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2581 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2641 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2701 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     2761 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2821 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2881 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2941 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3001 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3061 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3121 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3181 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     3241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3361 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtggccatt
     3421 aatccgagct cttacgcgcc cgcgtaaaac acaatcaagt atgagtcata agctgatgtc
     3481 atgttttgca cacggctcat aaccgaactg gctttacgag tagaattcta cttgtaacgc
     3541 acgatcagtg gatgatgtca tttgtttttc aaatcgagat gatgtcatgt tttgcacacg
     3601 gctcataaac tcgctttacg agtagaattc tacgtgtaac gcacgatcga ttgatgagtc
     3661 atttgttttg caatatgata tcatacaata tgactcattt gtttttcaaa accgaacttg
     3721 atttacgggt agaattctac ttgtaaagca caatcaaaaa gatgatgtca tttgtttttc
     3781 aaaactgaac tcgctttacg agtagaattc tacgtgtaaa acacaatcaa gaaatgatgt
     3841 catttgttat aaaaataaaa gctgatgtca tgttttgcac atggctcata actaaactcg
     3901 ctttacgggt agaattctac gcgcgtcgat gtctttgtga tgcgcgcgac atttttgtag
     3961 gttattgata aaatgaacgg atacgttgcc cgacattatc attaaatcct tggcgtagaa
     4021 tttgtcgggt ccattgtccg tgtgcgctag catgcccgta acggacctcg tacttttggc
     4081 ttcaaaggtt ttgcgcacag acaaaatgtg ccacacttgc agctctgcat gtgtgcgcgt
     4141 taccacaaat cccaacggcg cagtgtactt gttgtatgca aataaatctc gataaaggcg
     4201 cggcgcgcga atgcagctga tcacgtacgc tcctcgtgtt ccgttcaagg acggtgttat
     4261 cgacctcaga ttaatgttta tcggccgact gttttcgtat ccgctcacca aacgcgtttt
     4321 tgcattaaca ttgtatgtcg gcggatgttc tatatctaat ttgaataaat aaacgataac
     4381 cgcgttggtt ttagagggca taataaaaga aatattgtta tcgtgttcgc cattagggca
     4441 gtataaattg acgttcatgt tggatattgt ttcagttgca agttgacact ggcggcgaca
     4501 agatcgtgaa caaccaagtg accattaacc ctcgaccgcc ggagtataaa tagaggcgct
     4561 tcgtctacgg agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag
     4621 ctaagcaaat aaacaagcgc agctgaacaa gctaaacaat cggggtaccg ctagagtcga
     4681 cggtacgatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     4741 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4801 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     4861 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     4921 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4981 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     5041 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     5101 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     5161 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     5221 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     5281 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaggtgagt
     5341 agtacatgtg catacatctt aagttcactt gatctatagg aactgcgatt gcaacatcaa
     5401 attgtctgcg gcgtgagaac tgcgacccac aaaaatccca aaccgcggcc gtttatcacc
     5461 actttgtaca agaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
     5521 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
     5581 tggtcgacct gcagactggc tgtgtataag ggagcctgac atttatattc cccagaacat
     5641 caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc
     5701 gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc
     5761 gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc
     5821 cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta catccacaaa
     5881 cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt caccagcccc
     5941 tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca gccatccctt
     6001 cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc tgcatggttg
     6061 tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat agctgtcgct
     6121 gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat acttcgggta
     6181 tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc atactgttat
     6241 ctggctttta gtaagccgga tccacgcgtt tacgccccgc cctgccactc atcgcagtac
     6301 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     6361 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     6421 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     6481 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     6541 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     6601 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     6661 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     6721 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     6781 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     6841 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     6901 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgac
     6961 ggatcctaac tcaaaatcca cacattatac gagccggaag cataaagtgt aaagcctggg
     7021 gtgcctaatg cggccgccat agtgactgga tatgttgtgt tttacagtat tatgtagtct
     7081 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt
     7141 cagctttttt gtacaaactt gtgataaacg aattcttgaa gacgaaaggg cacaaacaaa
     7201 tagtgacacg aaacagatta ttctggtagc tgttctcgct atataagaca atttttgaga
     7261 tcatatcatg atcaagacat ctaaaggcat tcattttcga ctatattctt ttttacaaaa
     7321 aatataacaa ccagatattt taagctgatc ctagatgcac aaaaaataaa taaaagtata
     7381 aacctacttc gtaggatact tcggggtact ttttgttcgg ggttagatga gcataacgct
     7441 tgtagttgat atttgagatc ccctatcatt gcaggaccca aacgagaagc gtgatcacat
     7501 ggtcctcctc gagttcgtga ccgccgccgg catcaccctg ggcatggacg agctgtacaa
     7561 gtaagtatag tgccttgacc gagctcgccc actaagcgtc gcgccacttc aacgctcgat
     7621 gggagcgtca ttggtgggcg gggtaaccgt cgaaatcagt gtttacgctt ccaatcgcaa
     7681 caaaaaattc actgcaacac tgaaaagcat acgaaaacga tgaagattgt acgagaaacc
     7741 ataaagtatt ttatccacaa agacacgtat agcagaaaag ccaagttaac tcggcgataa
     7801 gttgtgtaca caagaataaa atcggccaga ttcagtgttg tcagaaataa gaaaacccca
     7861 ctatgttttt ctttgccttt tctttctccc agcgatcatt catttcgtgg tgaaagaacg
     7921 gggtcattgc acggagtttc gactgcggga aagcagagct gccgttcact tcgtctataa
     7981 ttagcgcttt ctattttccc cgattcgggc cgctgctgcg cttttccgcc tgctgtttgt
     8041 ggcaagtgta gcagcaggct gtgcacgcag tgtggcatgc acttggcttt ccaccgttgg
     8101 tatcgattct ctgggacgat gagtcattcc tttcggggcc acagcataat cgttgccagc
     8161 tcaccgaaat ggtgacttca tttcttaact gccgtcaagc atgcgattgt acata
//
