LOCUS       sequence-431358-        4389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of vdh.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4389)
  AUTHORS   Ruhl IA, Woodworth SP, Haugen SJ, Alt HM, Beckham GT, Johnson CW
  TITLE     Production of Vanillin From Ferulic Acid by Pseudomonas putida 
            KT2440 Using Metabolic Engineering and In Situ Product Recovery.
  JOURNAL   Microb Biotechnol. 2025 May;18(5):e70152. doi: 
            10.1111/1751-7915.70152.
  PUBMED    40410945
REFERENCE   2  (bases 1 to 4389)
  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: "Microb 
            Biotechnol."; date: "2025-05"; volume: "18(5):e70152. doi"; pages: "
            10.1111/1751-7915.70152"
COMMENT     Sequence Label: pIAR001
FEATURES             Location/Qualifiers
     source          1..4389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(821..1615)
                     /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(932..951)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1542..1561
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1696..1713
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1697..1713
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(3161..3749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3241..3260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3782..4389,1..727,728..814))
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 ggtagccatc ttcagttcca gtgtttgctt caaatactaa gtatttgtgg cctttatctt
       61 ctacgtagtg aggatctctc agcgtatggt tgtcgcctga gctgtagttg ccttcatcga
      121 tgaactgctg tacattttga tacgtttttc cgtcaccgtc aaagattgat ttataatcct
      181 ctacaccgtt gatgttcaaa gagctgtctg atgctgatac gttaacttgt gcagttgtca
      241 gtgtttgttt gccgtaatgt ttaccggaga aatcagtgta gaataaacgg atttttccgt
      301 cagatgtaaa tgtggctgaa cctgaccatt cttgtgtttg gtcttttagg atagaatcat
      361 ttgcatcgaa tttgtcgctg tctttaaaga cgcggccagc gtttttccag ctgtcaatag
      421 aagtttcgcc gactttttga tagaacatgt aaatcgatgt gtcatccgca tttttaggat
      481 ctccggctaa tgcaaagacg atgtggtagc cgtgatagtt tgcgacagtg ccgtcagcgt
      541 tttgtaatgg ccagctgtcc caaacgtcca ggccttttgc agaagagata tttttaattg
      601 tggacgaatc aaattcagaa acttgatatt tttcattttt ttgctgttca gggatttgca
      661 gcatatcatg gcgtgtaata tgggaaatgc cgtatgtttc cttatatggc ttttggttcg
      721 tttctttcgc aaacgcttga gttgcgcctc ctgccagcag tgcggtagta aaggttaata
      781 ctgttgcttg ttttgcaaac tttttgatgt tcatcgtcgc tcagaagaac tcgtcaagaa
      841 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
      901 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
      961 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     1021 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     1081 gcatccgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     1141 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     1201 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     1261 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     1321 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     1381 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttggagtt
     1441 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     1501 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     1561 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     1621 acgatcctca tcctgtttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     1681 ttccggtcac gacgttgtaa aacgacggcc agtgccaagc ttaagcttat atgggggcga
     1741 cctgcgacct ggccacgcac gctgatcacg ccgaccgaat cgtccagcac cgcctcgaag
     1801 gcgcgctgga acggctggct gtcgctgacg aagaccactc cgggggtgag cacctcgcag
     1861 acatggcgca acttggcgaa gtcttgcgac aacagcgcgt aggccggcga caccgggcaa
     1921 taggcaatac cggcatacat ggcgccgagg gcgatttgca ggtgttcgat gtcgttgccg
     1981 gaaagcagcg ccagcgggcg ctcggcactg aggcccagtc ctagcaagtt ggcggcgatg
     2041 gtgcgcacat cggcgagcat ctgcacgtag ctgatcgaac gccaggcacc gtctgcctgg
     2101 cgtgccgcga tgaaagtggt gtccgggcgc acctgggccc aatgcaccag gcgctcgagc
     2161 aggcgcgccg gcagcggcgc cagtggctcg acaggttgca tgcgcagcac gtcgtcgacg
     2221 tgactgacct gcacctgggg atgcccgatg gccacctggc ggtagcgcgg acgttggcca
     2281 gggtcggtcg accctgagcg ggcttcgtta ttcacgcagg ttgctccttg ttcttgttgt
     2341 acatcctgcc gccgcccgat atgacggggc ggcggtcatc ttgccggagg tctgggcctg
     2401 ccgcggctag atgggatagt gagatatcgt tgtcgttata gagaaatacc cgcgggcttt
     2461 ggcgcggcac accgccgccc gcggtgcgac tcagcgcttg taggcctgca ggcctggctt
     2521 gatgctcttg tcgtcgagga actgcttcat gccctgctcg cggccgccgg tagtgtccag
     2581 caggcgcgac tggtcgagct tggcgtagag gtagtcctcg ttctgttccc aggtcagctc
     2641 gcggcaacgc ttgaagccat tcttcgcggc acgcagcacc accgggttct tttccagcag
     2701 gttcagcgcc aactcgcggg tggtttcacg cagctcggcc agcggcacac tgtcgttcac
     2761 caggcccatc tcggcagcct tgcgaccatc gaaggtcttg ccggtcatga tgtagtacag
     2821 cgactgacga tggccaacgg tatcggccat ggccttgctg accaggttac caggcgggat
     2881 gccccagttg atttccgaca ggccgaaggt cgcttcgttg gcgcagatcg ccaggtcgca
     2941 tgccaccagt gggctgaagc cgccgccgaa gcaccagccg ttgaccatgg cgatggtcgg
     3001 tttggcatac agacgcagca acttccattg ccattgcgag gcttcgcgac gaatcttttc
     3061 ctggaggatt tccgggccgg cgtccacctc gcggaagtac tccttcaggt ccatgccgga
     3121 attcgtaatc atgtcatagc tgtttcctgt gtgaggcgtt tttccatagg ctccgccccc
     3181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3601 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3721 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3781 attatttgtt aactgttaat tgtccttgtt caaggatgct gtctttgaca acagatgttt
     3841 tcttgccttt gatgttcagc aggaagctcg gcgcaaacgt tgattgtttg tctgcgtaga
     3901 atcctctgtt tgtcatatag cttgtaatca cgacattgtt tcctttcgct tgaggtacag
     3961 cgaagtgtga gtaagtaaag gttacatcgt taggatcaag atccattttt aacacaaggc
     4021 cagttttgtt cagcggcttg tatgggccag ttaaagaatt agaaacataa ccaagcatgt
     4081 aaatatcgtt agacgtaatg ccgtcaatcg tcatttttga tccgcgggag tcagtgaaca
     4141 ggtaccattt gccgttcatt ttaaagacgt tcgcgcgttc aatttcatct gttactgtgt
     4201 tagatgcaat cagcggtttc atcacttttt tcagtgtgta atcatcgttt agctcaatca
     4261 taccgagagc gccgtttgct aactcagccg tgcgtttttt atcgctttgc agaagttttt
     4321 gactttcttg acggaagaat gatgtgcttt tgccatagta tgctttgtta aataaagatt
     4381 cttcgcctt
//
