LOCUS       sequence-357824-        6235 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 6235)
  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 6235)
  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{Mhc-F4-678-GFP::KD-RC,attB}
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(124..126,127..129,130..642)
                     /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(169..190)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(430..449)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    877..901
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(999..1023)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    2272..2305
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2346..2415
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2634..2654)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2634..2652)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2673..2689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2673..2689)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2686..2708)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2697..2713
                     /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(2721..2727,2728..2745,2746..2751))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2766..2787
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2904..2921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3075..3663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3155..3174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3834..4625,4626..4694))
                     /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     4457..4476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4695..4799)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4826..5281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4913..4932)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5123..5144
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5407..5429
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5421..5438
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5422..5438
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5448..5467
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5448..5466
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aattcaaaca agcaaagtga acacgtcgct aagcgaaagc taagcaaata aacaagcgca
       61 gctgaacaag ctaaacaatc ggggtaccgc tagagtcgac ggtacgatcc accggtcgcc
      121 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      181 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      241 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      301 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      421 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      541 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
      601 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      721 cactacctga gcacccagtc cgccctgagc aaggtgagta gtacatgtgc atacatctta
      781 agttcacttg atctatagga actgcgattg caacatcaaa ttgtctgcgg cgtgagaact
      841 gcgacccaca aaaatcccaa accgcggccg tttatcacca ctttgtacaa gaaagctggg
      901 tcggcgcgcc cacccttaaa cgatatcaga catttgtctg ataatgcttc attatcagac
      961 aaatgtctga tatcgtttgg tgaagggggc ggccgcggag cctgcttttt tgtacaaact
     1021 tgtgataaac gaattcttga agacgaaagg gcacaaacaa atagtgacac gaaacagatt
     1081 attctggtag ctgttctcgc tatataagac aatttttgag atcatatcat gatcaagaca
     1141 tctaaaggca ttcattttcg actatattct tttttacaaa aaatataaca accagatatt
     1201 ttaagctgat cctagatgca caaaaaataa ataaaagtat aaacctactt cgtaggatac
     1261 ttcggggtac tttttgttcg gggttagatg agcataacgc ttgtagttga tatttgagat
     1321 cccctatcat tgcaggaccc aaacgagaag cgtgatcaca tggtcctcct cgagttcgtg
     1381 accgccgccg gcatcaccct gggcatggac gagctgtaca agtaagtata gtgccttgac
     1441 cgagctcgcc cactaagcgt cgcgccactt caacgctcga tgggagcgtc attggtgggc
     1501 ggggtaaccg tcgaaatcag tgtttacgct tccaatcgca acaaaaaatt cactgcaaca
     1561 ctgaaaagca tacgaaaacg atgaagattg tacgagaaac cataaagtat tttatccaca
     1621 aagacacgta tagcagaaaa gccaagttaa ctcggcgata agttgtgtac acaagaataa
     1681 aatcggccag attcagtgtt gtcagaaata agaaaacccc actatgtttt tctttgcctt
     1741 ttctttctcc cagcgatcat tcatttcgtg gtgaaagaac ggggtcattg cacggagttt
     1801 cgactgcggg aaagcagagc tgccgttcac ttcgtctata attagcgctt tctattttcc
     1861 ccgattcggg ccgctgctgc gcttttccgc ctgctgtttg tggcaagtgt agcagcaggc
     1921 tgtgcacgca gtgtggcatg cacttggctt tccaccgttg gtatcgattc tctgggacga
     1981 tgagtcattc ctttcggggc cacagcataa tcgttgccag ctcaccgaaa tggtgacttc
     2041 atttcttaac tgccgtcaag catgcgattg tacatacata catatttata tatgtacata
     2101 tttatgtgac tatggtaggt cgatataata gcaatcaacg caagcaaatg tgtcagtcct
     2161 gcttacagga acgattctat ttagtaattt tcgttgtata aagtaattat gtatgtatgt
     2221 aagccccata aatctgaaac aattaggcaa aaccatgcga agctctctag tataacttcg
     2281 tataatgtat gctatacgaa gttctcgagg gggggcccgg taccgtcgac gatgtaggtc
     2341 acggtctcga agccgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
     2401 tccacctcac ccatctggtc catcatgatg aacgggtcga ggtggcggta gttgatcccg
     2461 gcgaacgcgc ggcgcgccgg gaagccctcg ccctcgaaac cgctgggcgc ggtggtcacg
     2521 gtgagcacgg gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag cgtcagcggg
     2581 ttctcgacgg tcacggcggg catgtcgaca ctagtggtac ccagcttttg ttccctttag
     2641 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2701 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2761 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2821 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2881 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2941 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3001 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3061 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3121 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3181 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3241 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3301 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3361 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3421 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3481 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3541 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3601 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3661 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3721 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3781 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3841 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3901 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3961 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4021 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4081 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4141 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4201 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4261 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4321 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4381 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4441 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4501 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4561 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4621 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4681 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4741 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4801 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4861 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4921 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4981 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5041 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5101 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5161 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5221 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5281 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5341 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     5401 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     5461 tatagggcga attggagctc caccgcggtg gccattaata tactcgaatt cgttgtcggc
     5521 tcatatacat gggcgaaata atttcgaata tgttttaaaa ataaccaaag acattagaaa
     5581 gagatcgcca atacttatac attatgtcta tgtgtgccat gtggtagcat gagccaaaaa
     5641 gcttctcgaa attacgaatt acatatagac gaatatgtat gtgactttag tttcgaaata
     5701 atttcgagaa ttttaaaaat aacgcattcg ttaaaagttc gcgtcaattc gaatcgaatt
     5761 ttcgattgtc gatttagtgt ggattgtcga aaatcgttcc gccttcgaag tttactgaaa
     5821 ggaatcattg cgatctcgtg aattgcttgt atgagaacac gccaccatat cgagatacct
     5881 gttcaaaata tcgaaatcgc gttccaaacg gtagtagtta ccagttgagt gagttgtggt
     5941 gacagtttaa cctctctaca tttgtaataa taagcattgt tttggcgtat caatagtctg
     6001 tagatttttt cacaaagaaa cctggagaaa tgtacacaaa tatctaaatg aaaattggcg
     6061 ttacgtaaca aataccaacg ccccgaaaat ccgtatatat gcaaagcatt ctgatatata
     6121 tatatataag tggtatacat atacatattc gtatcggatc cgtgatctat atttaggatt
     6181 aaccctcgac cgccggagta taaatagagg cgcttcgtct acggagcgac aattc
//
