LOCUS       sequence-335634-        3853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G418 sulfate (geneticin) resistance marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3853)
  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 3853)
  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: pNPTII
FEATURES             Location/Qualifiers
     source          1..3853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(154..742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(234..253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(913..1704,1705..1773))
                     /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     1536..1555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1774..1878)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1956..2411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2093..2114)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2305..2324
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2537..2559
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2551..2568
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2552..2568
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2575..2594
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2575..2593
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2624..2633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2628..2637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3569..3587)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3570..3587)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3605..3621)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3605..3621)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3618..3640)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3629..3645
                     /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(3653..3659,3660..3677,3678..3683))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3698..3719
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3836..3853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
       61 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1021 accatctggc cccagtgctg caatgatacc gcgcgaccca cgctcaccgg ctccagattt
     1081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac
     1921 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     1981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2161 tcagggcgct ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2401 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2461 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2521 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     2581 gactcactat agggcgaatt gggcccgacg tcgcatgctc ccggccgcca tggcggccgc
     2641 gggaattcga ttgggcgatg agtggtctcg aagcttagaa gaactcgtcc agcagacggt
     2701 agaaggcaat acgctggcta tctggggctg caatgccgta gagcaccagg aatcggtcgg
     2761 cccactcgcc tcccagctcc tcggcgatat ctcgggtggc cagggcgata tcctggtaac
     2821 ggtcggccac gcccaggcgt ccacaatcaa tgaagccact gaagcgtccg ttctccacca
     2881 taatgttggg caggcaggcg tcgccgtggg tcaccaccag gtcctcgcca tcgggcatac
     2941 gggccttcag acgggcaaac agctcggctg gagccaggcc ctgttgctcc tcatccaggt
     3001 catcctggtc caccagtcca gcctccatac gggttcgggc acgctcaata cggtgcttgg
     3061 cctggtggtc aaaggggcag gtggctgggt ccagggtgtg caggcgacgc atagcatcgg
     3121 ccataatgga caccttctcg gctggggcca gatggctgga cagcaggtcc tggcctggca
     3181 cctcgcccag cagcagccaa tcgcgtccag cttcggtcac cacatccagc acggcagcac
     3241 acggcactcc tgttgtggcc agccagctca gacgagctgc ctcatcctgg agctcgttca
     3301 gggctccact caggtcggtc ttgacgaaca gcacggggcg tccctgggcc gacagacgga
     3361 acacggcagc atcggagcag ccgatggtct gctgggccca atcgtagccg aacagacgct
     3421 ccacccaggc agctggcgag ccagcgtgca gtccatcctg ctcaatcatt cgagaccact
     3481 catcgcccaa tcactagtga attcgcggcc gcctgcaggt cgaccatatg ggagagctcc
     3541 caacgcgttg gatgcatagc ttgagtattc tatagtgtca cctaaatagc ttggcgtaat
     3601 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3661 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3721 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3781 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3841 tcactgactc gct
//
