LOCUS       sequence-335040-        5004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GB2.0 compatible FlyLight Enhancer element R70B04 in enhancer 
            grammar.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5004)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 5004)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pR70B04
FEATURES             Location/Qualifiers
     source          1..5004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(110..129)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(111..129)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(136..153)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(136..152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      294..321
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      347..802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(434..453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     644..665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        880..984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(985..1053,1054..1845)
                     /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     complement(1203..1222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2016..2604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2505..2524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2758..2775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2790..2861)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2932..2948
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2932..2948
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2964..2982
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2964..2981
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tgccacgtca atcgttaagt tactggcgat gggctgtctg tcggactgac attccctctt
       61 cacctcccga gaccttgaga tcaatatcta aggtcctaag cggaattcac cctatagtga
      121 gtcgtattac aattcactgg ccgtcgtttt acattgagaa gagaaaagaa aaccgccgat
      181 cctgtccacc gcattactgc aaggtagtgg acaagaccgg cggtcttaag ttttttggct
      241 gaaagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
      301 catcctgacg gatggccttt ttgcgtttct acaatgttac gaatggacgc gccctgtagc
      361 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      421 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      481 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      541 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccagc gccctgatag
      601 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      661 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
      721 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      781 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
      841 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      901 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      961 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1021 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1081 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1141 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1201 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1261 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1321 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1381 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1441 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1501 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1561 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1621 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1681 tctggagccg gtgagcgtgg gtcgcgcggt atcattgcag cactggggcc agatggtaag
     1741 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1801 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1861 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1921 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2161 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2581 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2641 ttttgctggc cttttgctca catgttcttt cctgcgttat cccatgattc tgtggataac
     2701 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2761 gagtcagtga gcgaggaagc ggaaggtcag agagcgttca ccgacaaaca acagataaaa
     2821 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctgct cggtaccaaa
     2881 ttccagaaaa gaggcctccc gaaagggggg ccttttttcg ttttggtccg acaggaaaca
     2941 gctatgacca acggctctat ggtatttagg tgacactata gaacgaattc actagtgatt
     3001 gggcgatgag tggtctcggg gatataagag ggctgcctgt attaggacca agtttttgtt
     3061 tgtggcttat tacttctgat gagtctaaca atcttctagg gaaaatggga agcgctacgc
     3121 ttataagcca cattgcttta atgaattatt cattgacaaa taaaacgttg cagccggctg
     3181 aataattgta aaatattagt tttatatcct gtttgcaaaa tgcaatttgg ttgcataatg
     3241 aagtgtataa ctaattgtat gcatctcatg ttgttcaaga cgaaatcgtt tttttttcgc
     3301 agaaatatct cgacttttaa attgtgctta actcttaatc attgtcttta tttcacataa
     3361 gtaatatttt ctacacatcc agtaacctgt tttcttaata tgttaactgc tttaattcaa
     3421 atttggcacc caagctgaaa tgttactgca ttcagctcag ccaggggaaa tctgcaaaaa
     3481 ttgaacagag aaatcacgtt tagttaacat taacagctcg gctggcttac aatctgcctg
     3541 caaagtggta aaagaaaatg cagacggtgc ctagaatttt cctgaactgt aaagctgtcc
     3601 aaatcaaaat tgatatcaac tgcagctata taattcaata acatgggatc gggaataatt
     3661 ccgaaatgtt ccactcgagc aatcctcata agagaaaacc cgagtatgtt tatggacacg
     3721 ataatcatca gtaatcaaaa caaaataaat taaagcttta taagctaaca ttagctgtcg
     3781 ggcagccaga gaacataaat cgagacgaga gcccctagca aacagaaata aaatggcaga
     3841 aattattatt atctaaacaa aaaaaggggg gagctcatat gccatcgctc aaaagcactc
     3901 tgctatcgaa atactgaggt ggccaagggg ccgagggggc ttcagcagcc cttgtttcca
     3961 tcgctgcgca ctcttgtttt tgggttcaaa ttgaaaaaca tgaaaagaaa tatttccaat
     4021 caattggcaa agagaaacga ataaaaattc acacacagaa cagacaaaaa acccaggcac
     4081 acacaccgga ggaggcaaca agaaaaatgt tgttcattca ttaaatacaa aaattcatta
     4141 aacagacgag cgatcagaaa tattactgct gcagtccatt ccaaagtggc ctggggtctg
     4201 accggtggaa aagggaaaaa ataaaaataa aaatagttgt ttactttcgg aacagaggca
     4261 gcgacaacaa caagagaaaa acacaggggt aacaagctct cacagtgcga taaaatatgt
     4321 ttttttaatg caggtccgtg ttcaataaaa gtgccacaaa tccaaataga agctgctgca
     4381 ggaaagttcc gaacaaaaac atttatccag ttcgagcatt tgataccttt ttggtatgaa
     4441 ctcaggccct atcactgaac gtcgtcctgc aaaaggatat accctctatt accctgaaat
     4501 gaccagtgaa aggcgggaaa gtgaatcatg caaaacgaat gaaagtgctt cgaacagcag
     4561 tagaacttga tttgcattcc atttacaagc tgttcataga taataattaa taatatgatt
     4621 taaatgtcta ttctaagcga cattggggga tttgctttaa taatacaaaa aactaatagg
     4681 tttcgcaaga aaaccaaatt aaacatttca gactttccat tagtatccat acctaaaaat
     4741 tgtaacgcag gcctttacat atcattgacc cttctaaaag cgaacggcaa gaaaccgact
     4801 tggcattggt gaagtgaatt tattgttcga ctaatagatg atgactgcta tatactcgca
     4861 ttcggagtgc cttggaaaag tgtcgagtaa tttgtggaaa tggtcaaggc gatgtgtatg
     4921 agcagagtgg gcaactggcc atttcccgtg gcagaccgca tttatcactc ctggcctctt
     4981 cgattttttt gaacgaattg ataa
//
