LOCUS       sequence-357826-        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-RC,attB}
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     510..529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(748..852)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      879..1334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(966..985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1176..1197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1460..1482
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1474..1491
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1475..1491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1501..1520
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1501..1519
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        1575..2057
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        2061..2652
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    3091..3115
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3132..3165)
                     /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(3186..3210)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4904..4937
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4978..5047
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(5266..5286)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5266..5284)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5305..5321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5305..5321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5318..5340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5329..5345
                     /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(5353..5359,5360..5377,5378..5383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5398..5419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5536..5553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5707..6295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5787..5806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6466..6579,1..678,679..747))
                     /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"
ORIGIN
        1 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
       61 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
      121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
      181 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
      241 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
      301 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
      361 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
      421 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
      481 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
      541 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
      601 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
      661 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
      721 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
      781 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
      841 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
      901 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      961 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1021 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1081 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1141 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1201 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1261 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1321 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     1381 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     1441 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     1501 taatacgact cactataggg cgaattggag ctccaccgcg gtggcggccg ctttaatccg
     1561 agctcttacg cgcccgcgta aaacacaatc aagtatgagt cataagctga tgtcatgttt
     1621 tgcacacggc tcataaccga actggcttta cgagtagaat tctacttgta acgcacgatc
     1681 agtggatgat gtcatttgtt tttcaaatcg agatgatgtc atgttttgca cacggctcat
     1741 aaactcgctt tacgagtaga attctacgtg taacgcacga tcgattgatg agtcatttgt
     1801 tttgcaatat gatatcatac aatatgactc atttgttttt caaaaccgaa cttgatttac
     1861 gggtagaatt ctacttgtaa agcacaatca aaaagatgat gtcatttgtt tttcaaaact
     1921 gaactcgctt tacgagtaga attctacgtg taaaacacaa tcaagaaatg atgtcatttg
     1981 ttataaaaat aaaagctgat gtcatgtttt gcacatggct cataactaaa ctcgctttac
     2041 gggtagaatt ctacgcgcgt cgatgtcttt gtgatgcgcg cgacattttt gtaggttatt
     2101 gataaaatga acggatacgt tgcccgacat tatcattaaa tccttggcgt agaatttgtc
     2161 gggtccattg tccgtgtgcg ctagcatgcc cgtaacggac ctcgtacttt tggcttcaaa
     2221 ggttttgcgc acagacaaaa tgtgccacac ttgcagctct gcatgtgtgc gcgttaccac
     2281 aaatcccaac ggcgcagtgt acttgttgta tgcaaataaa tctcgataaa ggcgcggcgc
     2341 gcgaatgcag ctgatcacgt acgctcctcg tgttccgttc aaggacggtg ttatcgacct
     2401 cagattaatg tttatcggcc gactgttttc gtatccgctc accaaacgcg tttttgcatt
     2461 aacattgtat gtcggcggat gttctatatc taatttgaat aaataaacga taaccgcgtt
     2521 ggttttagag ggcataataa aagaaatatt gttatcgtgt tcgccattag ggcagtataa
     2581 attgacgttc atgttggata ttgtttcagt tgcaagttga cactggcggc gacaagatcg
     2641 tgaacaacca agtgaccatt aaaaccctcg accgccggag tataaataga ggcgcttcgt
     2701 ctacggagcg acaattcaat tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa
     2761 gcaaataaac aagcgcagct gaacaagcta aacaatcggg cggccgcact agagccggtc
     2821 gccaccatga ggtcttccaa gaatgttatc aaggagttca tgaggtttaa ggttcgcatg
     2881 gaaggaacgg tcaatgggca cgagtttgaa atagaaggcg aaggagaggg gaggccatac
     2941 gaaggccaca ataccgtaaa gcttaaggtg agtagtacat gtgcatacat cttaagttca
     3001 cttgatctat aggaactgcg attgcaacat caaattgtct gcggcgtgag aactgcgacc
     3061 cacaaaaatc ccaaaccgcg gccgtttatc accactttgt acaagaaagc tgggtcggcg
     3121 cgcccaccct tgaagttcct atactttcta gagaatagga acttcggtga agggggcggc
     3181 cgcggagcct gcttttttgt acaaacttgt gataaacgaa ttcttgaaga cgaaagggca
     3241 caaacaaata gtgacacgaa acagattatt ctggtagctg ttctcgctat ataagacaat
     3301 ttttgagatc atatcatgat caagacatct aaaggcattc attttcgact atattctttt
     3361 ttacaaaaaa tataacaacc agatatttta agctgatcct agatgcacaa aaaataaata
     3421 aaagtataaa cctacttcgt aggatacttc ggggtacttt ttgttcgggg ttagatgagc
     3481 ataacgcttg tagttgatat ttgagatccc ctatcattgc aggtaaccaa ggggggacct
     3541 ttgccatttg cttgggatat tttgtcacca caatttcagt atggaagcaa ggtatatgtc
     3601 aagcaccctg ccgacatacc agactataaa aagctgtcat ttcctgaagg atttaaatgg
     3661 gaaagggtca tgaactttga agacggtggc gtcgttactg taacccagga ttccagtttg
     3721 caggatggct gtttcatcta caaggtcaag ttcattggcg tgaactttcc ttccgatgga
     3781 cctgttatgc aaaagaagac aatgggctgg gaagccagca ctgagcgttt gtatcctcgt
     3841 gatggcgtgt tgaaaggaga gattcataag gctctgaagc tgaaagacgg tggtcattac
     3901 ctagttgaat tcaaaagtat ttacatggca aagaagcctg tgcagctacc agggtactac
     3961 tatgttgact ccaaactgga tataacaagc cacaacgaag actatacaat cgttgagcag
     4021 tatgaaagaa ccgagggacg ccaccatctg ttcctttagc ggccatcgaa ttcgagctcg
     4081 cccactaagc gtcgcgccac ttcaacgctc gatgggagcg tcattggtgg gcggggtaac
     4141 cgtcgaaatc agtgtttacg cttccaatcg caacaaaaaa ttcactgcaa cactgaaaag
     4201 catacgaaaa cgatgaagat tgtacgagaa accataaagt attttatcca caaagacacg
     4261 tatagcagaa aagccaagtt aactcggcga taagttgtgt acacaagaat aaaatcggcc
     4321 agattcagtg ttgtcagaaa taagaaaacc ccactatgtt tttctttgcc ttttctttct
     4381 cccagcgatc attcatttcg tggtgaaaga acggggtcat tgcacggagt ttcgactgcg
     4441 ggaaagcaga gctgccgttc acttcgtcta taattagcgc tttctatttt ccccgattcg
     4501 ggccgctgct gcgcttttcc gcctgctgtt tgtggcaagt gtagcagcag gctgtgcacg
     4561 cagtgtggca tgcacttggc tttccaccgt tggtatcgat tctctgggac gatgagtcat
     4621 tcctttcggg gccacagcat aatcgttgcc agctcaccga aatggtgact tcatttctta
     4681 actgccgtca agcatgcgat tgtacataca tacatattta tatatgtaca tatttatgtg
     4741 actatggtag gtcgatataa tagcaatcaa cgcaagcaaa tgtgtcagtc ctgcttacag
     4801 gaacgattct atttagtaat tttcgttgta taaagtaatt atgtatgtat gtaagcccca
     4861 taaatctgaa acaattaggc aaaaccatgc gaagctctct agtataactt cgtataatgt
     4921 atgctatacg aagttctcga gggggggccc ggtaccgtcg acgatgtagg tcacggtctc
     4981 gaagccgcgg tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actccacctc
     5041 acccatctgg tccatcatga tgaacgggtc gaggtggcgg tagttgatcc cggcgaacgc
     5101 gcggcgcgcc gggaagccct cgccctcgaa accgctgggc gcggtggtca cggtgagcac
     5161 gggacgtgcg acggcgtcgg cgggtgcgga tacgcggggc agcgtcagcg ggttctcgac
     5221 ggtcacggcg ggcatgtcga cactagtggt acccagcttt tgttcccttt agtgagggtt
     5281 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6541 cgtcgtgtag ataactacga tacgggaggg cttaccatc
//
