LOCUS       sequence-357878-        8164 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 8164)
  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 8164)
  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::GW,attB}
FEATURES             Location/Qualifiers
     source          1..8164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1663..1696
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1737..1806
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2025..2045)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2025..2043)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2064..2080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2064..2080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2077..2099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2088..2104
                     /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(2112..2118,2119..2136,2137..2142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2157..2178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2295..2312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2466..3054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2546..2565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3225..4016,4017..4085))
                     /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     3848..3867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4086..4190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4217..4672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4304..4323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4514..4535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4798..4820
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4812..4829
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4813..4829
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4839..4858
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4839..4857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        4913..5395
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        5399..5990
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    6429..6553
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6594..6899)
                     /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(6696..6715)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(7241..7900)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7815..7834
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7954..7960,7961..7978,7979..7984))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(8009..8133)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aaacaaatag tgacacgaaa cagattattc tggtagctgt tctcgctata taagacaatt
       61 tttgagatca tatcatgatc aagacatcta aaggcattca ttttcgacta tattcttttt
      121 tacaaaaaat ataacaacca gatattttaa gctgatccta gatgcacaaa aaataaataa
      181 aagtataaac ctacttcgta ggatacttcg gggtactttt tgttcggggt tagatgagca
      241 taacgcttgt agttgatatt tgagatcccc tatcattgca ggtaaccaag gggggacctt
      301 tgccatttgc ttgggatatt ttgtcaccac aatttcagta tggaagcaag gtatatgtca
      361 agcaccctgc cgacatacca gactataaaa agctgtcatt tcctgaagga tttaaatggg
      421 aaagggtcat gaactttgaa gacggtggcg tcgttactgt aacccaggat tccagtttgc
      481 aggatggctg tttcatctac aaggtcaagt tcattggcgt gaactttcct tccgatggac
      541 ctgttatgca aaagaagaca atgggctggg aagccagcac tgagcgtttg tatcctcgtg
      601 atggcgtgtt gaaaggagag attcataagg ctctgaagct gaaagacggt ggtcattacc
      661 tagttgaatt caaaagtatt tacatggcaa agaagcctgt gcagctacca gggtactact
      721 atgttgactc caaactggat ataacaagcc acaacgaaga ctatacaatc gttgagcagt
      781 atgaaagaac cgagggacgc caccatctgt tcctttagcg gccatcgaat tcgagctcgc
      841 ccactaagcg tcgcgccact tcaacgctcg atgggagcgt cattggtggg cggggtaacc
      901 gtcgaaatca gtgtttacgc ttccaatcgc aacaaaaaat tcactgcaac actgaaaagc
      961 atacgaaaac gatgaagatt gtacgagaaa ccataaagta ttttatccac aaagacacgt
     1021 atagcagaaa agccaagtta actcggcgat aagttgtgta cacaagaata aaatcggcca
     1081 gattcagtgt tgtcagaaat aagaaaaccc cactatgttt ttctttgcct tttctttctc
     1141 ccagcgatca ttcatttcgt ggtgaaagaa cggggtcatt gcacggagtt tcgactgcgg
     1201 gaaagcagag ctgccgttca cttcgtctat aattagcgct ttctattttc cccgattcgg
     1261 gccgctgctg cgcttttccg cctgctgttt gtggcaagtg tagcagcagg ctgtgcacgc
     1321 agtgtggcat gcacttggct ttccaccgtt ggtatcgatt ctctgggacg atgagtcatt
     1381 cctttcgggg ccacagcata atcgttgcca gctcaccgaa atggtgactt catttcttaa
     1441 ctgccgtcaa gcatgcgatt gtacatacat acatatttat atatgtacat atttatgtga
     1501 ctatggtagg tcgatataat agcaatcaac gcaagcaaat gtgtcagtcc tgcttacagg
     1561 aacgattcta tttagtaatt ttcgttgtat aaagtaatta tgtatgtatg taagccccat
     1621 aaatctgaaa caattaggca aaaccatgcg aagctctcta gtataacttc gtataatgta
     1681 tgctatacga agttctcgag ggggggcccg gtaccgtcga cgatgtaggt cacggtctcg
     1741 aagccgcggt gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta ctccacctca
     1801 cccatctggt ccatcatgat gaacgggtcg aggtggcggt agttgatccc ggcgaacgcg
     1861 cggcgcgccg ggaagccctc gccctcgaaa ccgctgggcg cggtggtcac ggtgagcacg
     1921 ggacgtgcga cggcgtcggc gggtgcggat acgcggggca gcgtcagcgg gttctcgacg
     1981 gtcacggcgg gcatgtcgac actagtggta cccagctttt gttcccttta gtgagggtta
     2041 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2101 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2161 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2641 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2881 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4201 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4621 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc
     4681 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4741 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     4801 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     4861 aattggagct ccaccgcggt ggcggccgct ttaatccgag ctcttacgcg cccgcgtaaa
     4921 acacaatcaa gtatgagtca taagctgatg tcatgttttg cacacggctc ataaccgaac
     4981 tggctttacg agtagaattc tacttgtaac gcacgatcag tggatgatgt catttgtttt
     5041 tcaaatcgag atgatgtcat gttttgcaca cggctcataa actcgcttta cgagtagaat
     5101 tctacgtgta acgcacgatc gattgatgag tcatttgttt tgcaatatga tatcatacaa
     5161 tatgactcat ttgtttttca aaaccgaact tgatttacgg gtagaattct acttgtaaag
     5221 cacaatcaaa aagatgatgt catttgtttt tcaaaactga actcgcttta cgagtagaat
     5281 tctacgtgta aaacacaatc aagaaatgat gtcatttgtt ataaaaataa aagctgatgt
     5341 catgttttgc acatggctca taactaaact cgctttacgg gtagaattct acgcgcgtcg
     5401 atgtctttgt gatgcgcgcg acatttttgt aggttattga taaaatgaac ggatacgttg
     5461 cccgacatta tcattaaatc cttggcgtag aatttgtcgg gtccattgtc cgtgtgcgct
     5521 agcatgcccg taacggacct cgtacttttg gcttcaaagg ttttgcgcac agacaaaatg
     5581 tgccacactt gcagctctgc atgtgtgcgc gttaccacaa atcccaacgg cgcagtgtac
     5641 ttgttgtatg caaataaatc tcgataaagg cgcggcgcgc gaatgcagct gatcacgtac
     5701 gctcctcgtg ttccgttcaa ggacggtgtt atcgacctca gattaatgtt tatcggccga
     5761 ctgttttcgt atccgctcac caaacgcgtt tttgcattaa cattgtatgt cggcggatgt
     5821 tctatatcta atttgaataa ataaacgata accgcgttgg ttttagaggg cataataaaa
     5881 gaaatattgt tatcgtgttc gccattaggg cagtataaat tgacgttcat gttggatatt
     5941 gtttcagttg caagttgaca ctggcggcga caagatcgtg aacaaccaag tgaccattaa
     6001 aaccctcgac cgccggagta taaatagagg cgcttcgtct acggagcgac aattcaattc
     6061 aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc aaataaacaa gcgcagctga
     6121 acaagctaaa caatcgggcg gccgcactag agccggtcgc caccatgagg tcttccaaga
     6181 atgttatcaa ggagttcatg aggtttaagg ttcgcatgga aggaacggtc aatgggcacg
     6241 agtttgaaat agaaggcgaa ggagagggga ggccatacga aggccacaat accgtaaagc
     6301 ttaaggtgag tagtacatgt gcatacatct taagttcact tgatctatag gaactgcgat
     6361 tgcaacatca aattgtctgc ggcgtgagaa ctgcgaccca caaaaatccc aaaccgcggc
     6421 cgtttatcac cactttgtac aagaaagctg aacgagaaac gtaaaatgat ataaatatca
     6481 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     6541 tccagtcact atggtcgacc tgcagactgg ctgtgtataa gggagcctga catttatatt
     6601 ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct gagatcagcc
     6661 acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat catgcgccag
     6721 ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc tgacagcaga
     6781 cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat atcactctgt
     6841 acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt aaactgcatt
     6901 tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc gggcgacctc
     6961 agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac gggcttcatt
     7021 ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt gagcaactga
     7081 tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct ctttttgaca
     7141 tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg cgcaaatacg
     7201 catactgtta tctggctttt agtaagccgg atccacgcgt ttacgccccg ccctgccact
     7261 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     7321 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     7381 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     7441 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     7501 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     7561 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     7621 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     7681 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     7741 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     7801 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     7861 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     7921 aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa gcataaagtg
     7981 taaagcctgg ggtgcctaat gcggccgcca tagtgactgg atatgttgtg ttttacagta
     8041 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     8101 cgtttctcgt tcagcttttt tgtacaaact tgtgataaac gaattcttga agacgaaagg
     8161 gcac
//
