LOCUS       sequence-335039-        4825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GB2.0 compatible FlyLight Enhancer element R20A02 in enhancer 
            grammar.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4825)
  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 4825)
  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: pR20A02
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(209..228)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(210..228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(235..252)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(235..251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      393..420
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      446..901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(533..552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     743..764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        979..1083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1084..1152,1153..1944)
                     /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(1302..1321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2115..2703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2604..2623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2857..2874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2889..2960)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3031..3047
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3031..3047
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3063..3081
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3063..3080
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttcttcatat gagttacgca gaagtttatc aagtgcgaaa agtatttgca aagttgccat
       61 ttttccagtt tgtacttctt taagatacac atacttacgt gggtggcatc atggccgtaa
      121 aaccttacag aaaatccatt tgcggactgc ccttaaaagt ggtccccgag accttgagat
      181 caatatctaa ggtcctaagc ggaattcacc ctatagtgag tcgtattaca attcactggc
      241 cgtcgtttta cattgagaag agaaaagaaa accgccgatc ctgtccaccg cattactgca
      301 aggtagtgga caagaccggc ggtcttaagt tttttggctg aaagggtggc gggcaggacg
      361 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
      421 tgcgtttcta caatgttacg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      481 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      541 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      601 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      661 tagggtgatg gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg
      721 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      781 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      841 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
      901 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc
      961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1801 tcgcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2761 atgttctttc ctgcgttatc ccatgattct gtggataacc gtattaccgc ctttgagtga
     2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2881 gaaggtcaga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     2941 tgagcctttc gttttatttg atgcctgctc ggtaccaaat tccagaaaag aggcctcccg
     3001 aaaggggggc cttttttcgt tttggtccga caggaaacag ctatgaccaa cggctctatg
     3061 gtatttaggt gacactatag aacgaattca ctagtgattg ggcgatgagt ggtctcggga
     3121 gtacgcgacc tacagtacct tgagccctgt ttatctagca gatatatttc ggtgctcgcc
     3181 tttcgcttga gctactttat ctgcaattct gcaaccaata ttggcgctta actgcaggta
     3241 atttcttacc ccatttgatt cattttgagc tgcttacgcc gtcatggaat gtaaattcgg
     3301 tctcttaaca tataaatttt cactttcctt cgacacaagc cccaacccac caatggagct
     3361 ccacgagcag cgctttgatg gtatttcacc gcacagctgc caaatattta ttcgcattta
     3421 aaaagtattt tatgttgcat tttcccataa gcattctcct gggcactttg gctgctgctg
     3481 ccacgtgagt ccaaatacca gtgctcggtg ataagacatg cattcgacct cgccgaaggt
     3541 cctcgttgcc tccatccatc tgctgctgtt cccgtgtcct caaaaatgct ttggcgttgg
     3601 gttttatgcg agctgctcct tgggagtgtt tgttttaatc cacccccgac caagtcgccc
     3661 agatcttgcc tgaattgttt gtatgacaaa gttagcattt tgtaataaat tcgcttcaaa
     3721 aaatatttga atgtttatgg tcggtattca aattaaatac tatgctgcgc gaaacaagta
     3781 aagccgaaca ttaaacgagg taccaataaa atgtattttt tgaaaatttg cctttgaaat
     3841 ttttgcgccc gttgaaattt tcattactat aatggaaaag aataattttt taacaaaaaa
     3901 aaaaaaggaa aagtaaatga aacgtattac cttaaggaaa acaaggacca ggaaacgttc
     3961 caacagcccc ttcaactgca ctttctcttg tttgatttct cggctcaatg gattcactgc
     4021 aaacggccaa gatgtgtggt cttcaaactg aaatgccacg aaattagcag agttcttacc
     4081 aattccagca ggcaattaaa ggcacttcag ttgctgaacc aactgctagg gtaaataaca
     4141 caatgctaca tacacgttgg cagcgagagc tcaaagtaag gctttgtttt atcgaggaga
     4201 aacatttaca tttacatttt gcaactggca aacagaaatt caatgcaaaa taagagcaat
     4261 cgtgcgggtt cagggggcaa gtggcactgt ttgtgttgag caaaagttgt ggcaacaatg
     4321 ctcataaaac ataaacaaac aaacaaataa acaaacaaac gtcattgcgc ccggcagttt
     4381 atcaaccagc caatggaatg ggtggcactg ggtggctctg gtgggttgca taatacatgt
     4441 gtgcggagtg gttttagagg cggattccag cgggggaatg cccatcgcac cgcaccgcac
     4501 cgacctgaca atgcaatcaa gcgaaagtga cagagcggca ggagctgagc cactgacatc
     4561 gatatgtcga caactggtaa ctgctgcaca tgatgacggc accggggtat ttggattttg
     4621 tggctggcgg atgtggcaag ctggatggct ggatacacgg gggcacacaa caaaatggct
     4681 gcgccaaagt atttagccaa actttgacac aagttttcct ttctcgcatt ttttgtgctg
     4741 ggattccagc cattggcata gttgttggtc ttcgtttgat ttgaatattt tgtcactcga
     4801 ctgacgcact tggctttcgg ctcca
//
