LOCUS       sequence-431360-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of PP_1948.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4498)
  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 4498)
  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: pIAR003
FEATURES             Location/Qualifiers
     source          1..4498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(35..829)
                     /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(146..165)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     756..775
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     910..927
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     911..927
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2200..2211)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2456..2472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2456..2472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(2484..3072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2564..2583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3105..4439,4440..4498,1..28))
                     /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 cttgttttgc aaactttttg atgttcatcg tcgctcagaa gaactcgtca agaaggcgat
       61 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
      121 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
      181 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
      241 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatcc
      301 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
      361 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
      421 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
      481 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
      541 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
      601 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgg agttcattca
      661 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
      721 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
      781 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
      841 ctcatcctgt ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
      901 tcacgacgtt gtaaaacgac ggccagtgcc aagcttgact gattacccag ggagtactcg
      961 acaccggtgg cacgctcagc ctcaataacg attttttgga cgagcgcgcc cgtcaacaca
     1021 tgcaggtttg ggcgcttcag tacaggctga aggaatgcag tggccgccga gtgccgccgc
     1081 ccccctttaa tggtgaattg caggaagccg gtgccttcag ggcttggagc gttgagatcg
     1141 tccagacgag gaattcccgc ttcgacacaa gactcaatga acgcctgaga ggacggatgt
     1201 ttgaataccg ggtctgtgac ccaaagctcg ccatcgcggc ctcggaatgc ttcatcaccc
     1261 tcctcccgat gttcaaattt cttgtagaaa ggcaggacat cgtcccaacc ccacccagcg
     1321 catcctaggg atacccaatg atcgtagtcc tctctatggc ctcgaagata agccatccca
     1381 ttgatcgcac tcgatccacc cagtgctttt ccacgaggca catagatacg ccgccctgcc
     1441 aagctaggat cgggttcaga ttggtacgcc caattcgtcg gcccaggcag gatgacccgt
     1501 gacacccctg ctggcatcga agcccacagt ccctttggcc gtgaacccgc ttccagcagt
     1561 agcactgaat gctctgggtt ggcagaaagc cggtttgcca aaacacagcc ggcagatccc
     1621 gctccaatga ttatgtagtc gtacaccggc tgctcctcgc tcactcacaa cagctcgcac
     1681 ccgaagatat cgcacttttg attaatcgat taaaattttt ccaccacggg ttaccacaac
     1741 accatcgacg ctgatgcagg gctcccgaat tacaccttca agtcgaagct tcgccttaca
     1801 agtaccaccc agggctgttg tatcaccgag gccaatgtgt gaagttgcgt acattttttt
     1861 atcggtacgc atggagcccg tttgtttaac ttgcggattg agcccaatcg cgatttcagc
     1921 cggacaatta aagctttcag ggtcgccgtg cttagcgaaa atctcgcgcc acatagctgc
     1981 ttgcttgcca ccttcaatag acacggccat gccattctta acggttaccg taataggctc
     2041 cgctagggca cccaccgagt gagcggtcat gtcaaaaaca atcgtgccct cgccggttcc
     2101 ttcgaccggg cacacgtgtg tttcgccatc cggaaatgcg cagccactac cgccacctgg
     2161 ccgcataacg acaggtagcc cagtgatggc acgcccaaca cgaccttcaa tactggcggt
     2221 caaatgagta ccgtttggac aagtaacggt gacctgcttg ccgtttgtga aaagatcagc
     2281 aatcttcctg ccgaggatat tcagcgcatg gtaatcagca cccgctggac catccttatc
     2341 caacatggcc gtggtgactt cctccatgag caagaaacgg cgctcatgct caatcatcag
     2401 atcggtaatg ggggcatgtg ccagtgctgt actcgtcagg tgaattcgta atcatgtcat
     2461 agctgtttcc tgtgtgaggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2641 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2881 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3061 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgattattt gttaactgtt
     3121 aattgtcctt gttcaaggat gctgtctttg acaacagatg ttttcttgcc tttgatgttc
     3181 agcaggaagc tcggcgcaaa cgttgattgt ttgtctgcgt agaatcctct gtttgtcata
     3241 tagcttgtaa tcacgacatt gtttcctttc gcttgaggta cagcgaagtg tgagtaagta
     3301 aaggttacat cgttaggatc aagatccatt tttaacacaa ggccagtttt gttcagcggc
     3361 ttgtatgggc cagttaaaga attagaaaca taaccaagca tgtaaatatc gttagacgta
     3421 atgccgtcaa tcgtcatttt tgatccgcgg gagtcagtga acaggtacca tttgccgttc
     3481 attttaaaga cgttcgcgcg ttcaatttca tctgttactg tgttagatgc aatcagcggt
     3541 ttcatcactt ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt
     3601 gctaactcag ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag
     3661 aatgatgtgc ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca
     3721 tcttcagttc cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag
     3781 tgaggatctc tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc
     3841 tgtacatttt gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg
     3901 ttgatgttca aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt
     3961 ttgccgtaat gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta
     4021 aatgtggctg aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg
     4081 aatttgtcgc tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg
     4141 ccgacttttt gatagaacat gtaaatcgat gtgtcatccg catttttagg atctccggct
     4201 aatgcaaaga cgatgtggta gccgtgatag tttgcgacag tgccgtcagc gttttgtaat
     4261 ggccagctgt cccaaacgtc caggcctttt gcagaagaga tatttttaat tgtggacgaa
     4321 tcaaattcag aaacttgata tttttcattt ttttgctgtt cagggatttg cagcatatca
     4381 tggcgtgtaa tatgggaaat gccgtatgtt tccttatatg gcttttggtt cgtttctttc
     4441 gcaaacgctt gagttgcgcc tcctgccagc agtgcggtag taaaggttaa tactgttg
//
