LOCUS       sequence-333403-        2616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generic suicide vector for knockouts and integrations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2616)
  AUTHORS   Cook TB, Jacobson TB, Venkataraman MV, Hofstetter H, Amador-Noguez 
            D, Thomas MG, Pfleger BF
  TITLE     Stepwise genetic engineering of Pseudomonas putida enables robust 
            heterologous production of prodigiosin and glidobactin A.
  JOURNAL   Metab Eng. 2021 Jun 24;67:112-124. doi: 10.1016/j.ymben.2021.06.004.
  PUBMED    34175462
REFERENCE   2  (bases 1 to 2616)
  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: "Metab 
            Eng."; date: "2021-06-24"; volume: "67:112-124. doi"; pages: " 
            10.1016/j.ymben.2021.06.004"
COMMENT     Sequence Label: pNVLTv2
FEATURES             Location/Qualifiers
     source          1..2616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             256..1050
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(310..329)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     920..939
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1442..1458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1442..1458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1507..1524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1641..2186)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1721..1740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2262..2281
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            2409..2517
                     /note="incP origin of transfer"
     primer_bind     2547..2565
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct
       61 tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg
      121 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg
      181 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga
      241 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
      301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
      361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
      421 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
      481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
      541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
      601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
      661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
      721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
      781 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
      841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
      901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
      961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     1021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     1081 aagcgacgcc caacctgtca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     1141 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1201 tgctggagtt cttcgcccac cccgggctcg atcccctcgc gagttggttc agctgctgcc
     1261 tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc atccaggaaa
     1321 ccagcagcgg ctatccgcgc atccatgccc ccgaactgca ggagtgggga ggcacgatgg
     1381 ccgctttggt cccggatcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat
     1441 ggtcatagct gtttcctgtg tgaaattgcg tcagcagaat atgtgataca ggatatattc
     1501 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     1561 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     1621 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     1681 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1741 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     1801 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     1861 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     1921 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     1981 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     2041 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     2101 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     2161 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     2221 aggctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     2281 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     2341 atctgctcat gtttgacagc ttatcgctca gtggaacgaa aactcgcatg tatccaacgg
     2401 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     2461 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag
     2521 tgcttggatt ctcaccaata tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2581 gtttcggtga tgacggtgaa aacctctgag atgaat
//
