LOCUS       sequence-335645-        3626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  universal Vector Domesticator vector v2 used in the integration 
            (domestication) of new vector backbone parts in the GB2.0 cloning 
            system. Uses blue-white screening method.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3626)
  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 3626)
  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: pVD2
FEATURES             Location/Qualifiers
     source          1..3626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(138..155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(309..897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(389..408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1068..1859,1860..1928))
                     /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     1691..1710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1929..2033)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2111..2566)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2248..2269)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2460..2479
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2707..2723
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2730..2749
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2730..2748
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2779..2788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2783..2792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2917..2936
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3145..3161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3162..3218
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3231..3247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3255..3271
                     /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(3279..3285,3286..3303,3304..3309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3324..3345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(3497..3515)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3498..3515)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3533..3549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3557..3573
                     /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(3581..3587,3588..3605,3606..3611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(3626,1..21)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
       61 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
      121 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
      181 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      241 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      481 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      721 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      781 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1081 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1141 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1201 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1261 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1321 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1381 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1441 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1501 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1561 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1621 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1681 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1741 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1801 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1861 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1921 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2041 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2101 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2161 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2221 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2281 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2341 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2401 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2461 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2521 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     2581 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     2641 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     2701 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc
     2761 cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgattggg aagactctga
     2821 gacccacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2881 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2941 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3001 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggcg caggcctctt
     3061 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     3121 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc
     3181 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg
     3241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3301 tagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgg gtctcagagt cttcccaatc
     3421 actagtgaat tcgcggccgc ctgcaggtcg accatatggg agagctccca acgcgttgga
     3481 tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc
     3541 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3601 taaagtgtaa agcctggggt gcctaa
//
