LOCUS       sequence-357880-        7145 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 7145)
  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 7145)
  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-dsRed::GW,attB}
FEATURES             Location/Qualifiers
     source          1..7145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(145..936,937..1005))
                     /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     768..787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1006..1110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1137..1592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1224..1243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1434..1455
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1718..1740
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1732..1749
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1733..1749
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1759..1778
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1759..1777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1810..1826
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1810..1826
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2330..2454
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2495..2800)
                     /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(2597..2616)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(3142..3801)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3716..3735
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3855..3861,3862..3879,3880..3885))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3910..4034)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    5728..5761
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5802..5871
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(6090..6110)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6090..6108)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6129..6145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6129..6145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6142..6164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6153..6169
                     /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(6177..6183,6184..6201,6202..6207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6222..6243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6360..6377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6531..7119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6611..6630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
       61 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
      241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
      481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
      781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     1141 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1201 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1261 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1321 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1381 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1441 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1501 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1561 gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt
     1621 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     1681 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     1741 acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct ccaccgcggt
     1801 ggcggccgct ctagaactag tggatccccc ggggatctaa ttcaattaga gactaattca
     1861 attagagcta attcaattag gatccaagct tatcgatttc gaaccctcga ccgccggagt
     1921 ataaatagag gcgcttcgtc tacggagcga caattcaatt caaacaagca aagtgaacac
     1981 gtcgctaagc gaaagctaag caaataaaca agcgcagctg aacaagctaa acaatcgggc
     2041 ggccgcacta gagccggtcg ccaccatgag gtcttccaag aatgttatca aggagttcat
     2101 gaggtttaag gttcgcatgg aaggaacggt caatgggcac gagtttgaaa tagaaggcga
     2161 aggagagggg aggccatacg aaggccacaa taccgtaaag cttaaggtga gtagtacatg
     2221 tgcatacatc ttaagttcac ttgatctata ggaactgcga ttgcaacatc aaattgtctg
     2281 cggcgtgaga actgcgaccc acaaaaatcc caaaccgcgg ccgtttatca ccactttgta
     2341 caagaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     2401 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatggtcgac
     2461 ctgcagactg gctgtgtata agggagcctg acatttatat tccccagaac atcaggttaa
     2521 tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg
     2581 agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca
     2641 ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga
     2701 tcaccatccg tcgcccgggc gtgtcaataa tatcactctg tacatccaca aacagacgat
     2761 aacggctctc tcttttatag gtgtaaacct taaactgcat ttcaccagcc cctgttctcg
     2821 tcagcaaaag agccgttcat ttcaataaac cgggcgacct cagccatccc ttcctgattt
     2881 tccgctttcc agcgttcggc acgcagacga cgggcttcat tctgcatggt tgtgcttacc
     2941 agaccggaga tattgacatc atatatgcct tgagcaactg atagctgtcg ctgtcaactg
     3001 tcactgtaat acgctgcttc atagcatacc tctttttgac atacttcggg tatacatatc
     3061 agtatatatt cttataccgc aaaaatcagc gcgcaaatac gcatactgtt atctggcttt
     3121 tagtaagccg gatccacgcg tttacgcccc gccctgccac tcatcgcagt actgttgtaa
     3181 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     3241 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     3301 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     3361 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     3421 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     3481 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     3541 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
     3601 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     3661 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     3721 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     3781 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg acggatccta
     3841 actcaaaatc cacacattat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3901 tgcggccgcc atagtgactg gatatgttgt gttttacagt attatgtagt ctgtttttta
     3961 tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttt
     4021 ttgtacaaac ttgtgataaa cgaattcttg aagacgaaag ggcacaaaca aatagtgaca
     4081 cgaaacagat tattctggta gctgttctcg ctatataaga caatttttga gatcatatca
     4141 tgatcaagac atctaaaggc attcattttc gactatattc ttttttacaa aaaatataac
     4201 aaccagatat tttaagctga tcctagatgc acaaaaaata aataaaagta taaacctact
     4261 tcgtaggata cttcggggta ctttttgttc ggggttagat gagcataacg cttgtagttg
     4321 atatttgaga tcccctatca ttgcaggtaa ccaagggggg acctttgcca tttgcttggg
     4381 atattttgtc accacaattt cagtatggaa gcaaggtata tgtcaagcac cctgccgaca
     4441 taccagacta taaaaagctg tcatttcctg aaggatttaa atgggaaagg gtcatgaact
     4501 ttgaagacgg tggcgtcgtt actgtaaccc aggattccag tttgcaggat ggctgtttca
     4561 tctacaaggt caagttcatt ggcgtgaact ttccttccga tggacctgtt atgcaaaaga
     4621 agacaatggg ctgggaagcc agcactgagc gtttgtatcc tcgtgatggc gtgttgaaag
     4681 gagagattca taaggctctg aagctgaaag acggtggtca ttacctagtt gaattcaaaa
     4741 gtatttacat ggcaaagaag cctgtgcagc taccagggta ctactatgtt gactccaaac
     4801 tggatataac aagccacaac gaagactata caatcgttga gcagtatgaa agaaccgagg
     4861 gacgccacca tctgttcctt tagcggccat cgaattcgag ctcgcccact aagcgtcgcg
     4921 ccacttcaac gctcgatggg agcgtcattg gtgggcgggg taaccgtcga aatcagtgtt
     4981 tacgcttcca atcgcaacaa aaaattcact gcaacactga aaagcatacg aaaacgatga
     5041 agattgtacg agaaaccata aagtatttta tccacaaaga cacgtatagc agaaaagcca
     5101 agttaactcg gcgataagtt gtgtacacaa gaataaaatc ggccagattc agtgttgtca
     5161 gaaataagaa aaccccacta tgtttttctt tgccttttct ttctcccagc gatcattcat
     5221 ttcgtggtga aagaacgggg tcattgcacg gagtttcgac tgcgggaaag cagagctgcc
     5281 gttcacttcg tctataatta gcgctttcta ttttccccga ttcgggccgc tgctgcgctt
     5341 ttccgcctgc tgtttgtggc aagtgtagca gcaggctgtg cacgcagtgt ggcatgcact
     5401 tggctttcca ccgttggtat cgattctctg ggacgatgag tcattccttt cggggccaca
     5461 gcataatcgt tgccagctca ccgaaatggt gacttcattt cttaactgcc gtcaagcatg
     5521 cgattgtaca tacatacata tttatatatg tacatattta tgtgactatg gtaggtcgat
     5581 ataatagcaa tcaacgcaag caaatgtgtc agtcctgctt acaggaacga ttctatttag
     5641 taattttcgt tgtataaagt aattatgtat gtatgtaagc cccataaatc tgaaacaatt
     5701 aggcaaaacc atgcgaagct ctctagtata acttcgtata atgtatgcta tacgaagttc
     5761 tcgagggggg gcccggtacc gtcgacgatg taggtcacgg tctcgaagcc gcggtgcggg
     5821 tgccagggcg tgcccttggg ctccccgggc gcgtactcca cctcacccat ctggtccatc
     5881 atgatgaacg ggtcgaggtg gcggtagttg atcccggcga acgcgcggcg cgccgggaag
     5941 ccctcgccct cgaaaccgct gggcgcggtg gtcacggtga gcacgggacg tgcgacggcg
     6001 tcggcgggtg cggatacgcg gggcagcgtc agcgggttct cgacggtcac ggcgggcatg
     6061 tcgacactag tggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     6121 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6181 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6301 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6841 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6961 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7141 acggg
//
