LOCUS       sequence-357875-        7740 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 7740)
  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 7740)
  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-DsRed::GW,attB}
FEATURES             Location/Qualifiers
     source          1..7740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(72..96)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    1790..1823
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1864..1933
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2152..2172)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2152..2170)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2191..2207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2191..2207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2204..2226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2215..2231
                     /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(2239..2245,2246..2263,2264..2269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2284..2305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2422..2439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2593..3181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2673..2692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3352..4143,4144..4212))
                     /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     3975..3994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4213..4317)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4344..4799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4431..4450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4641..4662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4925..4947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4939..4956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4940..4956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4966..4985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4966..4984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6132..6256
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6297..6602)
                     /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(6399..6418)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6944..7603)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7518..7537
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7657..7663,7664..7681,7682..7687))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
ORIGIN
        1 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
       61 ttacgtttct cgttcagctt ttttgtacaa acttgtgata aacgaattct tgaagacgaa
      121 agggcacaaa caaatagtga cacgaaacag attattctgg tagctgttct cgctatataa
      181 gacaattttt gagatcatat catgatcaag acatctaaag gcattcattt tcgactatat
      241 tcttttttac aaaaaatata acaaccagat attttaagct gatcctagat gcacaaaaaa
      301 taaataaaag tataaaccta cttcgtagga tacttcgggg tactttttgt tcggggttag
      361 atgagcataa cgcttgtagt tgatatttga gatcccctat cattgcaggt aaccaagggg
      421 ggacctttgc catttgcttg ggatattttg tcaccacaat ttcagtatgg aagcaaggta
      481 tatgtcaagc accctgccga cataccagac tataaaaagc tgtcatttcc tgaaggattt
      541 aaatgggaaa gggtcatgaa ctttgaagac ggtggcgtcg ttactgtaac ccaggattcc
      601 agtttgcagg atggctgttt catctacaag gtcaagttca ttggcgtgaa ctttccttcc
      661 gatggacctg ttatgcaaaa gaagacaatg ggctgggaag ccagcactga gcgtttgtat
      721 cctcgtgatg gcgtgttgaa aggagagatt cataaggctc tgaagctgaa agacggtggt
      781 cattacctag ttgaattcaa aagtatttac atggcaaaga agcctgtgca gctaccaggg
      841 tactactatg ttgactccaa actggatata acaagccaca acgaagacta tacaatcgtt
      901 gagcagtatg aaagaaccga gggacgccac catctgttcc tttagcggcc atcgaattcg
      961 agctcgccca ctaagcgtcg cgccacttca acgctcgatg ggagcgtcat tggtgggcgg
     1021 ggtaaccgtc gaaatcagtg tttacgcttc caatcgcaac aaaaaattca ctgcaacact
     1081 gaaaagcata cgaaaacgat gaagattgta cgagaaacca taaagtattt tatccacaaa
     1141 gacacgtata gcagaaaagc caagttaact cggcgataag ttgtgtacac aagaataaaa
     1201 tcggccagat tcagtgttgt cagaaataag aaaaccccac tatgtttttc tttgcctttt
     1261 ctttctccca gcgatcattc atttcgtggt gaaagaacgg ggtcattgca cggagtttcg
     1321 actgcgggaa agcagagctg ccgttcactt cgtctataat tagcgctttc tattttcccc
     1381 gattcgggcc gctgctgcgc ttttccgcct gctgtttgtg gcaagtgtag cagcaggctg
     1441 tgcacgcagt gtggcatgca cttggctttc caccgttggt atcgattctc tgggacgatg
     1501 agtcattcct ttcggggcca cagcataatc gttgccagct caccgaaatg gtgacttcat
     1561 ttcttaactg ccgtcaagca tgcgattgta catacataca tatttatata tgtacatatt
     1621 tatgtgacta tggtaggtcg atataatagc aatcaacgca agcaaatgtg tcagtcctgc
     1681 ttacaggaac gattctattt agtaattttc gttgtataaa gtaattatgt atgtatgtaa
     1741 gccccataaa tctgaaacaa ttaggcaaaa ccatgcgaag ctctctagta taacttcgta
     1801 taatgtatgc tatacgaagt tctcgagggg gggcccggta ccgtcgacga tgtaggtcac
     1861 ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc
     1921 cacctcaccc atctggtcca tcatgatgaa cgggtcgagg tggcggtagt tgatcccggc
     1981 gaacgcgcgg cgcgccggga agccctcgcc ctcgaaaccg ctgggcgcgg tggtcacggt
     2041 gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg cggggcagcg tcagcgggtt
     2101 ctcgacggtc acggcgggca tgtcgacact agtggtaccc agcttttgtt ccctttagtg
     2161 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2221 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2281 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2341 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2401 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2461 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2521 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2581 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2641 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2701 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2761 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2821 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2881 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2941 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3001 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3061 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3121 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3181 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3241 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3301 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3361 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3421 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3481 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3541 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3601 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3661 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3721 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3781 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3841 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3901 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3961 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4021 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4081 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4141 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4201 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4261 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4321 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4381 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4441 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4501 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4561 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4621 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4681 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4741 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     4801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     4921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     4981 tagggcgaat tggagctcca ccgcggtggc ggccgcttta atatactcga attcgttgtc
     5041 ggctcatata catgggcgaa ataatttcga atatgtttta aaaataacca aagacattag
     5101 aaagagatcg ccaatactta tacattatgt ctatgtgtgc catgtggtag catgagccaa
     5161 aaagcttctc gaaattacga attacatata gacgaatatg tatgtgactt tagtttcgaa
     5221 ataatttcga gaattttaaa aataacgcat tcgttaaaag ttcgcgtcaa ttcgaatcga
     5281 attttcgatt gtcgatttag tgtggattgt cgaaaatcgt tccgccttcg aagtttactg
     5341 aaaggaatca ttgcgatctc gtgaattgct tgtatgagaa cacgccacca tatcgagata
     5401 cctgttcaaa atatcgaaat cgcgttccaa acggtagtag ttaccagttg agtgagttgt
     5461 ggtgacagtt taacctctct acatttgtaa taataagcat tgttttggcg tatcaatagt
     5521 ctgtagattt tttcacaaag aaacctggag aaatgtacac aaatatctaa atgaaaattg
     5581 gcgttacgta acaaatacca acgccccgaa aatccgtata tatgcaaagc attctgatat
     5641 atatatataa gtggtataca tatacatatt cgtatcggat ccgtgatcta tatttaggat
     5701 taaaaccctc gaccgccgga gtataaatag aggcgcttcg tctacggagc gacaattcaa
     5761 ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc
     5821 tgaacaagct aaacaatcgg gcggccgcac tagagccggt cgccaccatg aggtcttcca
     5881 agaatgttat caaggagttc atgaggttta aggttcgcat ggaaggaacg gtcaatgggc
     5941 acgagtttga aatagaaggc gaaggagagg ggaggccata cgaaggccac aataccgtaa
     6001 agcttaaggt gagtagtaca tgtgcataca tcttaagttc acttgatcta taggaactgc
     6061 gattgcaaca tcaaattgtc tgcggcgtga gaactgcgac ccacaaaaat cccaaaccgc
     6121 ggccgtttat caccactttg tacaagaaag ctgaacgaga aacgtaaaat gatataaata
     6181 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
     6241 atatccagtc actatggtcg acctgcagac tggctgtgta taagggagcc tgacatttat
     6301 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca
     6361 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc
     6421 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc
     6481 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc
     6541 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
     6601 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac
     6661 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc
     6721 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac
     6781 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg
     6841 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat
     6901 acgcatactg ttatctggct tttagtaagc cggatccacg cgtttacgcc ccgccctgcc
     6961 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     7021 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     7081 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     7141 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     7201 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     7261 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     7321 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     7381 tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     7441 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     7501 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     7561 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     7621 ctgaaaatct cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa
     7681 gtgtaaagcc tggggtgcct aatgcggccg ccatagtgac tggatatgtt gtgttttaca
//
