LOCUS       sequence-357881-        7205 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 7205)
  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 7205)
  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{3P3-mTurquoise::GW,attB}
FEATURES             Location/Qualifiers
     source          1..7205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(3..122)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1368..1401
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1442..1511
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(1730..1750)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1730..1748)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1769..1785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1769..1785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1782..1804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1793..1809
                     /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(1817..1823,1824..1841,1842..1847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1862..1883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2000..2017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2171..2759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2251..2270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2930..3721,3722..3790))
                     /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     3553..3572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3791..3895)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3922..4377
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4009..4028)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4219..4240
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4503..4525
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4517..4534
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4518..4534
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4544..4563
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4544..4562
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4861..4870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    5623..5747
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(5788..6093)
                     /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(5890..5909)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6435..7094)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7009..7028
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7148..7154,7155..7172,7173..7178))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
ORIGIN
        1 agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa
       61 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttttt gtacaaactt
      121 gtgataaacg aattcttgaa gacgaaaggg cacaaacaaa tagtgacacg aaacagatta
      181 ttctggtagc tgttctcgct atataagaca atttttgaga tcatatcatg atcaagacat
      241 ctaaaggcat tcattttcga ctatattctt ttttacaaaa aatataacaa ccagatattt
      301 taagctgatc ctagatgcac aaaaaataaa taaaagtata aacctacttc gtaggatact
      361 tcggggtact ttttgttcgg ggttagatga gcataacgct tgtagttgat atttgagatc
      421 ccctatcatt gcagccaaac gagaagcgtg atcacatggt cctcctcgag ttcgtgaccg
      481 ccgccggcat caccctgggc atggacgagc tgtacaagta agtatagtgc cttgaccgag
      541 ctcgcccact aagcgtcgcg ccacttcaac gctcgatggg agcgtcattg gtgggcgggg
      601 taaccgtcga aatcagtgtt tacgcttcca atcgcaacaa aaaattcact gcaacactga
      661 aaagcatacg aaaacgatga agattgtacg agaaaccata aagtatttta tccacaaaga
      721 cacgtatagc agaaaagcca agttaactcg gcgataagtt gtgtacacaa gaataaaatc
      781 ggccagattc agtgttgtca gaaataagaa aaccccacta tgtttttctt tgccttttct
      841 ttctcccagc gatcattcat ttcgtggtga aagaacgggg tcattgcacg gagtttcgac
      901 tgcgggaaag cagagctgcc gttcacttcg tctataatta gcgctttcta ttttccccga
      961 ttcgggccgc tgctgcgctt ttccgcctgc tgtttgtggc aagtgtagca gcaggctgtg
     1021 cacgcagtgt ggcatgcact tggctttcca ccgttggtat cgattctctg ggacgatgag
     1081 tcattccttt cggggccaca gcataatcgt tgccagctca ccgaaatggt gacttcattt
     1141 cttaactgcc gtcaagcatg cgattgtaca tacatacata tttatatatg tacatattta
     1201 tgtgactatg gtaggtcgat ataatagcaa tcaacgcaag caaatgtgtc agtcctgctt
     1261 acaggaacga ttctatttag taattttcgt tgtataaagt aattatgtat gtatgtaagc
     1321 cccataaatc tgaaacaatt aggcaaaacc atgcgaagct ctctagtata acttcgtata
     1381 atgtatgcta tacgaagttc tcgagggggg gcccggtacc gtcgacgatg taggtcacgg
     1441 tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca
     1501 cctcacccat ctggtccatc atgatgaacg ggtcgaggtg gcggtagttg atcccggcga
     1561 acgcgcggcg cgccgggaag ccctcgccct cgaaaccgct gggcgcggtg gtcacggtga
     1621 gcacgggacg tgcgacggcg tcggcgggtg cggatacgcg gggcagcgtc agcgggttct
     1681 cgacggtcac ggcgggcatg tcgacactag tggtacccag cttttgttcc ctttagtgag
     1741 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1801 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1861 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1921 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1981 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2041 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2101 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2161 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2221 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2281 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2341 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2401 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2461 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2521 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2581 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2641 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2701 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2761 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2821 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2881 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2941 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3001 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3061 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3121 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3181 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3241 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3301 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3361 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3421 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3481 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3541 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3601 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3661 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3721 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3781 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3841 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3901 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3961 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4021 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4081 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4141 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4201 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4261 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4321 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca
     4381 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4441 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     4501 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     4561 gggcgaattg gagctccacc gcggtggcca tatgcaatgg ttaattcgag ctcgcccggg
     4621 gatctaattc aattagagac taattcaatt agagctaatt caattaggat ccaagcttat
     4681 cgatttcgaa ccctcgaccg ccggagtata aatagaggcg cttcgtctac ggagcgacaa
     4741 ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc
     4801 gcagctgaac aagctaaaca atcggggtac cgctagagtc gacggtacga tccaccggtc
     4861 gccaccatgg tgtccaaggg agaggagctg ttcaccggag tcgtgccaat cctcgtcgag
     4921 ctggatggtg acgtgaacgg ccataagttc tccgtgtccg gtgagggcga aggtgacgcc
     4981 acctacggaa agctgaccct gaagttcatc tgcaccaccg gtaagctgcc cgtcccctgg
     5041 cccaccctgg tcaccaccct gtcctgggga gtgcagtgct tcgctcgcta ccccgaccac
     5101 atgaagcagc acgacttctt caagtccgcc atgccagaag gatacgttca ggagcgcacc
     5161 atcttcttca aggacgatgg caactacaag acccgcgccg aggtgaagtt cgagggcgat
     5221 accctggtca accgtatcga gctgaagggc atcgacttca aggaggacgg caacatcttg
     5281 ggtcacaagc tggagtacaa ctacattagc gacaacgtgt acatcaccgc tgataagcag
     5341 aagaacggca tcaaggctaa cttcaagatc cgtcacaaca tcgaggatgg cggcgtccag
     5401 ctggctgatc actaccagca gaacaccccc attggtgatg gtcccgtgct gctgcccgat
     5461 aatcactacc tgtccaccca gtccaagctc tccaaggacg tgagtagtac atgtgcatac
     5521 atcttaagtt cacttgatct ataggaactg cgattgcaac atcaaattgt ctgcggcgtg
     5581 agaactgcga cccacaaaaa tcccaaaccg cggccgttta tcaccacttt gtacaagaaa
     5641 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     5701 aaacagacta cataatactg taaaacacaa catatccagt cactatggtc gacctgcaga
     5761 ctggctgtgt ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt
     5821 tttgatgtca ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg
     5881 cacactggcc atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg
     5941 gtaaagttca cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat
     6001 ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct
     6061 ctctctttta taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcagcaa
     6121 aagagccgtt catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt
     6181 tccagcgttc ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accagaccgg
     6241 agatattgac atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt
     6301 aatacgctgc ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat
     6361 attcttatac cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttttagtaag
     6421 ccggatccac gcgtttacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     6481 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     6541 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     6601 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     6661 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     6721 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     6781 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     6841 catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg
     6901 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     6961 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     7021 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     7081 atttttttct ccattttagc ttccttagct cctgaaaatc tcgacggatc ctaactcaaa
     7141 atccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgcggcc
     7201 gccat
//
