LOCUS       sequence-357885-        6579 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 6579)
  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 6579)
  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-DsRed::FRT,attB}
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    165..198
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    239..308
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(527..547)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(527..545)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(566..582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(566..582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(579..601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    590..606
                     /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(614..620,621..638,639..644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    659..680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(797..814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(968..1556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1048..1067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1727..2518,2519..2587))
                     /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     2350..2369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2588..2692)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2719..3174
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2806..2825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3016..3037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3300..3322
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3314..3331
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3315..3331
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3341..3360
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3341..3359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3415..3897
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        3901..4492
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    4931..4955
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4972..5005
                     /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(5026..5050)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gactatggta ggtcgatata atagcaatca acgcaagcaa atgtgtcagt cctgcttaca
       61 ggaacgattc tatttagtaa ttttcgttgt ataaagtaat tatgtatgta tgtaagcccc
      121 ataaatctga aacaattagg caaaaccatg cgaagctctc tagtataact tcgtataatg
      181 tatgctatac gaagttctcg agggggggcc cggtaccgtc gacgatgtag gtcacggtct
      241 cgaagccgcg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccacct
      301 cacccatctg gtccatcatg atgaacgggt cgaggtggcg gtagttgatc ccggcgaacg
      361 cgcggcgcgc cgggaagccc tcgccctcga aaccgctggg cgcggtggtc acggtgagca
      421 cgggacgtgc gacggcgtcg gcgggtgcgg atacgcgggg cagcgtcagc gggttctcga
      481 cggtcacggc gggcatgtcg acactagtgg tacccagctt ttgttccctt tagtgagggt
      541 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      601 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      661 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1381 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2101 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2701 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2761 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2821 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2881 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2941 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3001 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3061 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3121 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
     3181 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     3241 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     3301 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     3361 cgaattggag ctccaccgcg gtggcggccg ctttaatccg agctcttacg cgcccgcgta
     3421 aaacacaatc aagtatgagt cataagctga tgtcatgttt tgcacacggc tcataaccga
     3481 actggcttta cgagtagaat tctacttgta acgcacgatc agtggatgat gtcatttgtt
     3541 tttcaaatcg agatgatgtc atgttttgca cacggctcat aaactcgctt tacgagtaga
     3601 attctacgtg taacgcacga tcgattgatg agtcatttgt tttgcaatat gatatcatac
     3661 aatatgactc atttgttttt caaaaccgaa cttgatttac gggtagaatt ctacttgtaa
     3721 agcacaatca aaaagatgat gtcatttgtt tttcaaaact gaactcgctt tacgagtaga
     3781 attctacgtg taaaacacaa tcaagaaatg atgtcatttg ttataaaaat aaaagctgat
     3841 gtcatgtttt gcacatggct cataactaaa ctcgctttac gggtagaatt ctacgcgcgt
     3901 cgatgtcttt gtgatgcgcg cgacattttt gtaggttatt gataaaatga acggatacgt
     3961 tgcccgacat tatcattaaa tccttggcgt agaatttgtc gggtccattg tccgtgtgcg
     4021 ctagcatgcc cgtaacggac ctcgtacttt tggcttcaaa ggttttgcgc acagacaaaa
     4081 tgtgccacac ttgcagctct gcatgtgtgc gcgttaccac aaatcccaac ggcgcagtgt
     4141 acttgttgta tgcaaataaa tctcgataaa ggcgcggcgc gcgaatgcag ctgatcacgt
     4201 acgctcctcg tgttccgttc aaggacggtg ttatcgacct cagattaatg tttatcggcc
     4261 gactgttttc gtatccgctc accaaacgcg tttttgcatt aacattgtat gtcggcggat
     4321 gttctatatc taatttgaat aaataaacga taaccgcgtt ggttttagag ggcataataa
     4381 aagaaatatt gttatcgtgt tcgccattag ggcagtataa attgacgttc atgttggata
     4441 ttgtttcagt tgcaagttga cactggcggc gacaagatcg tgaacaacca agtgaccatt
     4501 aaaaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat
     4561 tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct
     4621 gaacaagcta aacaatcggg cggccgcact agagccggtc gccaccatga ggtcttccaa
     4681 gaatgttatc aaggagttca tgaggtttaa ggttcgcatg gaaggaacgg tcaatgggca
     4741 cgagtttgaa atagaaggcg aaggagaggg gaggccatac gaaggccaca ataccgtaaa
     4801 gcttaaggtg agtagtacat gtgcatacat cttaagttca cttgatctat aggaactgcg
     4861 attgcaacat caaattgtct gcggcgtgag aactgcgacc cacaaaaatc ccaaaccgcg
     4921 gccgtttatc accactttgt acaagaaagc tgggtcggcg cgcccaccct tgaagttcct
     4981 attctctaga aagtatagga acttcggtga agggggcggc cgcggagcct gcttttttgt
     5041 acaaacttgt gataaacgaa ttcttgaaga cgaaagggca caaacaaata gtgacacgaa
     5101 acagattatt ctggtagctg ttctcgctat ataagacaat ttttgagatc atatcatgat
     5161 caagacatct aaaggcattc attttcgact atattctttt ttacaaaaaa tataacaacc
     5221 agatatttta agctgatcct agatgcacaa aaaataaata aaagtataaa cctacttcgt
     5281 aggatacttc ggggtacttt ttgttcgggg ttagatgagc ataacgcttg tagttgatat
     5341 ttgagatccc ctatcattgc aggtaaccaa ggggggacct ttgccatttg cttgggatat
     5401 tttgtcacca caatttcagt atggaagcaa ggtatatgtc aagcaccctg ccgacatacc
     5461 agactataaa aagctgtcat ttcctgaagg atttaaatgg gaaagggtca tgaactttga
     5521 agacggtggc gtcgttactg taacccagga ttccagtttg caggatggct gtttcatcta
     5581 caaggtcaag ttcattggcg tgaactttcc ttccgatgga cctgttatgc aaaagaagac
     5641 aatgggctgg gaagccagca ctgagcgttt gtatcctcgt gatggcgtgt tgaaaggaga
     5701 gattcataag gctctgaagc tgaaagacgg tggtcattac ctagttgaat tcaaaagtat
     5761 ttacatggca aagaagcctg tgcagctacc agggtactac tatgttgact ccaaactgga
     5821 tataacaagc cacaacgaag actatacaat cgttgagcag tatgaaagaa ccgagggacg
     5881 ccaccatctg ttcctttagc ggccatcgaa ttcgagctcg cccactaagc gtcgcgccac
     5941 ttcaacgctc gatgggagcg tcattggtgg gcggggtaac cgtcgaaatc agtgtttacg
     6001 cttccaatcg caacaaaaaa ttcactgcaa cactgaaaag catacgaaaa cgatgaagat
     6061 tgtacgagaa accataaagt attttatcca caaagacacg tatagcagaa aagccaagtt
     6121 aactcggcga taagttgtgt acacaagaat aaaatcggcc agattcagtg ttgtcagaaa
     6181 taagaaaacc ccactatgtt tttctttgcc ttttctttct cccagcgatc attcatttcg
     6241 tggtgaaaga acggggtcat tgcacggagt ttcgactgcg ggaaagcaga gctgccgttc
     6301 acttcgtcta taattagcgc tttctatttt ccccgattcg ggccgctgct gcgcttttcc
     6361 gcctgctgtt tgtggcaagt gtagcagcag gctgtgcacg cagtgtggca tgcacttggc
     6421 tttccaccgt tggtatcgat tctctgggac gatgagtcat tcctttcggg gccacagcat
     6481 aatcgttgcc agctcaccga aatggtgact tcatttctta actgccgtca agcatgcgat
     6541 tgtacataca tacatattta tatatgtaca tatttatgt
//
