LOCUS       sequence-329254-        7298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G418 resistance vector with attB and eye marker. Contains an Alpha 
            GB cloning entry point for addition of custom assembly uses 
            blue-white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  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 7298)
  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: pG418R-GMRWhite
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3..21)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(28..45)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(28..44)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(37..59)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(253..272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    329..398
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     825..846
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     polyA_signal    4286..4420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4310..4329)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4364..4383
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4735..5323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4815..4834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5791..6582
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6637..7092
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6724..6743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6934..6955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7116..7163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7217..7235
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7217..7234
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    7242..7298
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
       61 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      121 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      181 atgacgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
      241 gtgcactctc agtacaatct gctctgatgc cgcatagggt ctctctccgg atcccaggga
      301 ggaattggtc gacgatgtag gtcactgtct cgaagccgcg gtgcgggtgc cagggcgtgc
      361 ccttgggctc cccgggcgcg tactccacct cacccatctg gtccatcatg atgcgctgtc
      421 aggagctaga atcccaaaac aaactggtta ttgtggtagg tcatttgttt ggcagaaaga
      481 aaactcgaga aatttctctg gccgttattc gttattctct cttttctttt tgggtcgctc
      541 cctctctgca ctaatgctct ctcactctgt cacacagtaa acggcatact gctctcgttg
      601 gttcgagaga gcgcgcctcg aatgttcgcg aaaagagcgc cggagtataa atagaggcgc
      661 ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa
      721 gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tctgcagtaa agtgcaagtt
      781 aaagtgaatc aattaaaagt aaccagcaac caagtaaatc aactgcaact actgaaatct
      841 gccaagaagt aattattgaa tacaagaaga gaacactcaa tactcgaatc catgtgggag
      901 tttattcttg acacagatat ttatgatata ataactgagt aagcttaaca taaggaggaa
      961 aacaaaatga ttgagcagga tggactgcac gctggctcgc cagctgcctg ggtggagcgt
     1021 ctgttcggct acgattgggc ccagcagacc atcggctgct ccgatgctgc cgtgttccgt
     1081 ctgtcggccc agggacgccc cgtgctgttc gtcaagaccg acctgagtgg agccctgaac
     1141 gagctccagg atgaggcagc tcgtctgagc tggctggcca caacaggagt gccgtgtgct
     1201 gccgtgctgg atgtggtgac cgaagctgga cgcgattggc tgctgctggg cgaggtgcca
     1261 ggccaggacc tgctgtccag ccatctggcc ccagccgaga aggtgtccat tatggccgat
     1321 gctatgcgtc gcctgcacac cctggaccca gccacctgcc cctttgacca ccaggccaag
     1381 caccgtattg agcgtgcccg aacccgtatg gaggctggac tggtggacca ggatgacctg
     1441 gatgaggagc accagggcct ggctccagcc gagctgtttg cccgtctgaa ggcccgtatg
     1501 cccgatggcg aggacctggt ggtgacccac ggcgacgcct gcctgcccaa cattatggtg
     1561 gagaacggac gcttcagtgg cttcattgat tgtggacgcc tgggcgtggc cgaccgttac
     1621 caggatatcg ccctggccac ccgagatatc gccgaggagc tgggaggcga gtgggccgac
     1681 cgattcctgg tgctctacgg cattgcagcc ccagatagcc agcgtattgc cttctaccgt
     1741 ctgctggacg agttcttcta agcttggcaa taaaaagaca gaataaaacg cacgggtgtt
     1801 gggtcgtttg ttccgctgtc aggagcgtac gactagcacc tgactgtcga gcccagtgga
     1861 aacccttgaa atgcctttaa gtcgagccca gtggaaaccc ttgaaatgcc tttaagtcga
     1921 gcccagtgga aacccttgaa atgcctttaa gtcgagccca gtggaaaccc ttgaaatgcc
     1981 tttaagtcga gcccagtgga aacccttgaa atgcctttaa gtcgagtagc agtcgaggta
     2041 ggtacgtcat gcccctcccg agctcgcccg gggatcgagc gcagcggtat aaaagggcgc
     2101 ggggtggctg agagcatcag ttgtgaatga atgttcgagc cgagcagacg tgccgctgcc
     2161 ttcgttaata tcctttgaat aagccaactt tgaatcacaa gacgcatacc aaaccaaatg
     2221 ggacaggagg accaggagct gctgatccgc ggaggctcca agcacccaag cgccgagcac
     2281 ctgaacaacg gcgatagcgg agccgccagc cagtcgtgca tcaatcaggg atttggccag
     2341 gccaagaatt acggaaccct gctgcccccc agccccccag aggatagcgg aagcggatcc
     2401 ggacagctgg ccgagaatct gacgtacgcc tggcataaca tggatatctt cggcgccgtg
     2461 aaccagccag gcagtggctg gcgtcagctg gtgaaccgca cccgcggact gttctgcaat
     2521 gagcgccata tcccagcccc acgcaagcac ctgctgaaga atgtgtgcgg agtggcctac
     2581 cccggcgagc tgctggccgt gatgggaagt agtggcgccg gaaagaccac gctgctgaac
     2641 gccctggcct tccgcagccc ccagggcatt caggtgtccc caagcggcat gcgcctgctg
     2701 aacggccagc ccgtggatgc caaggagatg caggcccgct gcgcctacgt gcagcaggac
     2761 gatctgttca tcggcagcct gacagcccgc gagcatctga tctttcaggc catggtccgc
     2821 atgccccgcc acctgacata ccgtcagcgc gtggcccgcg tggaccaggt catccaggag
     2881 ctgagcctga gcaagtgcca gcacaccatt attggcgtgc caggccgcgt gaagggcctg
     2941 agtggcggag agcgcaagcg cctggccttt gccagcgagg ccctgaccga tccccccctg
     3001 ctgatttgcg acgagccgac cagcggcctg gatagcttca ccgcccactc cgtggtgcag
     3061 gtgctgaaga agctgagcca gaagggaaag acggtcattc tgacgatcca ccagcccagc
     3121 tcggagctgt tcgagctgtt tgataagatt ctgctgatgg ccgagggacg cgtggccttc
     3181 ctgggaaccc caagcgaggc cgtggatttc ttcagctatg tgggcgccca gtgccccacg
     3241 aattacaacc ccgccgactt ctatgtgcaa gtgctggccg tggtgccagg acgcgagatt
     3301 gagagccgcg atcgcattgc caagatctgc gataacttcg ccatctccaa ggtggcccgc
     3361 gacatggagc agctgctggc cacgaagaac ctggagaagc ccctggagca gcccgagaac
     3421 ggctatacgt ataaggctac ctggtttatg cagtttcgcg ccgtgctgtg gcgcagctgg
     3481 ctgagtgtgc tgaaggagcc cctgctggtc aaggtccgcc tgatccagac cacgatggtg
     3541 gccattctga tcggcctgat cttcctggga cagcagctga cgcaggtcgg agtgatgaac
     3601 atcaatggag ccatttttct gtttctgacg aacatgacgt tccagaacgt gttcgccacc
     3661 atcaacgtgt tcaccagcga gctgcccgtg ttcatgcgcg aggcccgcag ccgcctgtac
     3721 cgctgcgata cctacttcct gggaaagacc attgccgagc tgccgctgtt cctgaccgtg
     3781 cccctggtgt ttaccgccat tgcctacccc atgattggcc tgcgtgccgg agtgctgcat
     3841 ttctttaatt gcctggccct ggtgaccctg gtggccaacg tgtcgaccag cttcggctac
     3901 ctgatcagct gcgcctccag cagcaccagc atggccctga gcgtgggacc ccccgtgatt
     3961 atccccttcc tgctgttcgg cggattcttt ctgaactccg gcagcgtgcc cgtgtacctg
     4021 aagtggctga gctacctgag ctggtttcgc tacgccaatg agggcctgct gatcaatcag
     4081 tgggccgatg tggagcccgg cgagatctcc tgcaccagca gcaacacgac ctgccccagc
     4141 agcggaaagg tcatcctgga gacactgaat ttcagtgccg ccgacctgcc actggattat
     4201 gtgggcctgg ccatcctgat tgtgtccttc cgcgtgctgg cctatctggc tctgcgcctg
     4261 cgcgctcgtc gcaaggagtg agcttgatcc agacatgata agatacattg atgagtttgg
     4321 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4381 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4441 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcagtatggc aagcgctcga
     4501 cgaattaaca agcttcgctc tatctcgctc tctatctctc tcgatcgctc gtgctaagct
     4561 taagaccgtg gagaacaagg agaaccgctg tcaggaggca tgcgtcgact ctagacccgg
     4621 gcagctgcgc ttgacacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5401 attatcaaaa aggatcttca cctagatcct tttgtgacca gccaggacag aaatgcctcg
     5461 acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga
     5521 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca
     5581 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
     5641 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
     5701 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     5761 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg
     5821 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     5881 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     5941 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     6001 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     6061 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     6121 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     6181 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     6241 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     6301 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     6361 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt agctgccgac
     6421 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     6481 tatcttggac aagaagaaga tcgtttggcc tcgcgcgcag atcagttgga agaatttgtc
     6541 cactacgtga aaggcgagat caccaaggta gtcggcaaat aatcactaga ccaatgttac
     6601 acatatatac tttagattga tttaaaactt catgttacgc gccctgtagc ggcgcattaa
     6661 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6721 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6781 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6841 aaaaacttga ttagggtgat ggttcacgta gtgggccagc gccctgatag acggtttttc
     6901 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6961 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7021 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     7081 cgcttacaat tttaacggat cagcgagaga cccctgttga caattaatca tcggcatagt
     7141 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgacc
     7201 atgattacgc caagctattt aggtgacact atagaatact caagcttgca tgcctgcagg
     7261 tcgactctag aggatccccg ggtaccgagc tcgaattc
//
