LOCUS       sequence-335234-        4486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GB2.0 compatible FlyLight Enhancer element R76H03in enhancer 
            grammar.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4486)
  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 4486)
  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: pR76H03
FEATURES             Location/Qualifiers
     source          1..4486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(321..340)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(322..340)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(347..364)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(347..363)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      505..532
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      558..1013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(645..664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     855..876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1091..1195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1196..1264,1265..2056)
                     /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(1414..1433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2227..2815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2716..2735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2969..2986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3001..3072)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3143..3159
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3143..3159
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3175..3193
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3175..3192
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3282..3299)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ttactaatta attaattaag aaatcgacgc aaataaataa caaaaaacta attcaatata
       61 tgcatatgct agatatagtg gagttgattc cattgcaaaa tcgccgtacg gagagatttg
      121 atgtttaatt tttgactttg tttttattac agattaattt ttttaagttt tgcttagcga
      181 ctaaattttg ttatgcgtat taaaattgct tgcttgagtg tgtgtgtgtg tgtgtgtttg
      241 gacatgcgtg tctatgtgag cgtgtggagt cgtactcccg agaccttgag atcaatatct
      301 aaggtcctaa gcggaattca ccctatagtg agtcgtatta caattcactg gccgtcgttt
      361 tacattgaga agagaaaaga aaaccgccga tcctgtccac cgcattactg caaggtagtg
      421 gacaagaccg gcggtcttaa gttttttggc tgaaagggtg gcgggcagga cgcccgccat
      481 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
      541 tacaatgtta cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      601 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
      781 tggttcacgt agtgggccag cgccctgata gacggttttt cgccctttga cgttggagtc
      841 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
      901 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      961 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     1021 cggtattttc tacttacgca tctgtgcggt atttcacacc gcatcaggtg gcacttttcg
     1081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1441 tgacgccggg caagagcaac taggtcgccg catacactat tctcagaatg acttggttga
     1501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcgcgcgg
     1921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaaggtca
     3001 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt
     3061 tcgttttatt tgatgcctgc tcggtaccaa attccagaaa agaggcctcc cgaaaggggg
     3121 gccttttttc gttttggtcc gacaggaaac agctatgacc aacggctcta tggtatttag
     3181 gtgacactat agaacgaatt cactagtgat tgggcgatga gtggtctcgg ggagtgacaa
     3241 atgtgcagga ctggcttttg gtagcggtgg ttgccatgtt gatgatggtg gtgatggtgc
     3301 cacttttgct gccgtggcca ggggcgccca ctccactcca tccccaaatc tgggtctctg
     3361 caaggttctg gttgggtgtg ggtgcgagtt cgagttcgtg ccctggtcct gctcctactc
     3421 ctgctcctgc tcctgctcct ggtgcggcca ttgcagttat gtcacatgtc tgacttaatt
     3481 cattcattta gacgacgctg acaattgtag gtgtcgggtg cgcgccctgt gtcatttcgt
     3541 ttttgtcaac gtcgtcgacg cagtgcacca aatgggccag gggagtggat ggaattgggt
     3601 ggcggcgatg ggatataatg gcttgcggac gaagccaaag atagagaata ctggccaccg
     3661 gagccagtgg agagccatta tggcaccgac attgtcaatg actggcgaat tattattgcc
     3721 tgacggaggc agttggacgc caacgaagtg agcgcacccc ctccaccccc acagccacgc
     3781 cccattgtcc tggttgccac tgccacattt atgtacatat gtatttagag gtgtacgtgt
     3841 atgtgtgtat gtaagtgtgc tccatcttgt tatgacagtt ggcaccaatt tccgcacacg
     3901 ttgcaggtgg cccatggccc gtgggcgttg gaaaattgcg aggggggcgg ggtctaccac
     3961 caaccatttg tcggccggtc aatcaaaagt tggcgacctt tagacttatt gcattacatg
     4021 gcgttatttc actgctgttc actaaattga ttgcacttta atgcttttca tacaaaatta
     4081 tgaataacag tatttaaacg tatgccttcg gttcatttgc attttatttc acgataaata
     4141 aattatattg aaattatttc ttgaatttct aacgcctttt caaaatagca gtacattgca
     4201 cggttgcatt gggcgccgag ttgccagtta gcggaaatgg ggatgccaga tgcatgtatt
     4261 taagtttcaa cttaaaaagt aaccgtgaaa ataaagacac gttaaaataa tgcaaccctt
     4321 cgcatattca gctttccgtt gtatattaac ccataaaaag taacaacata attgtatata
     4381 tgtatatatt tcatatgtat catacaaaat ttccatttta agatttagca actaaattgg
     4441 attgagtttt tgtttcgttt tgaagtgcac aaattaaaaa aaaaga
//
