LOCUS       sequence-431367-        4499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of ior_operon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4499)
  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 4499)
  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: pIAR014
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..72
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     56..72
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1602..1618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1602..1618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(1630..2218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1710..1729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2251..3585,3586..3672))
                     /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"
     CDS             complement(3679..4473)
                     /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(3790..3809)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4400..4419
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gggcttccca accttaccag agggcgcccc agctggcaat tccggtcacg acgttgtaaa
       61 acgacggcca gtgccaagct tccggaatac cccccgtacc tgtggcgcga cccgctgaac
      121 cctcagcgac tgatcggtgc caatgccgac ctgctcaaat acctgggcaa gcagttgggg
      181 ctggagatag aggtggttta cgccgggccg tggtcacgag cccaggaaga agtgcgcacc
      241 gggcggatcg acttgatggc cgggtatttc ctgaccgaag cgcgccggca gcatgtggac
      301 ttcattgccc cgccgttttt gcgcacaccc agcgtcatct gggtgcgcca ggacaatgcc
      361 tttgcctacc agcaatgggc cgacctcaaa ggccgcaagg gtggcactct ggtcaacaat
      421 agccacggcc aacagttcga tgaatacgcc caggccagtc tcaccctcga agcggtgccc
      481 actgccaggc aggcgttcga gaagctgatg cacaagcgca gcgattacgt ggtgtacgag
      541 cagtatcctg gcatggcgct ggcgcgcaca ttgggcatcg cagccactgt gcaggttttg
      601 cagccaccgg tgtccagcga agggttgtac ctggcaatct cacatgagtc gccttgcaac
      661 cggccactgc tgcgtgaggc gctggcgcgg gagatggcga agatcgttgg cggggccttg
      721 cccgagcagt ggttgaaaca gaatctggag cggtggcagt gacagagtgg ctcatgtgct
      781 gcgcggtctt tgttgggggc agcctcgcgt cggtacatgt gccactccaa tgatatcggt
      841 ttcacctgcc cgtcacggtt gttgtcgctg tgttttatct gcatggggaa tacgttgtcc
      901 cccggtgcag aaacgctagc acaggaggat gaccgctggt ctggtggcgg gggtgacttc
      961 agaggatcag gcaagtaatc cggcggaata cgcaagccag cttcgaaaaa tcaagcctgc
     1021 agtcggcgtt gaacaggact tgcgcaacga aaacccggat aatgccggcc aatttcgttt
     1081 tgcacgctta aatcacggta atcccgcatg gaaatcaagg tcaattttct cgacaatctc
     1141 cgtctcgaag ccaaattcga tgacttcacg gtgatcgccg accaacctat ccgctacaaa
     1201 ggcgatggtt cggccccagg cccgttcgac tatttcctgg cgtcttcggc cttgtgcgcg
     1261 gcttacttcg tcaagctgta ctgccagacc cgcgatattc ccaccgaaaa tatccgcctg
     1321 tcgcagaaca acatcgtcga cccggaaaac cgctacaacc agatcttcaa gatccaggtc
     1381 gaactgcccg aagacatctc cgacaaggac cgtcaaggca tcctgcgctc catcgaccgc
     1441 tgcacggtaa aaaaggtggt gcagaccggc cctgagttcg tgatcgaagt ggtcgacaac
     1501 ctcgatgccg atgcccaagg gttgctgatg ccggtgtcgg acaccagcac ctatatcccc
     1561 ggcaaggacc tgccgctgga gcagaccgaa ttcgtaatca tgtcatagct gtttcctgtg
     1621 tgaggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2041 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2221 agatcctttg atcttttcta cggggtctga ttatttgtta actgttaatt gtccttgttc
     2281 aaggatgctg tctttgacaa cagatgtttt cttgcctttg atgttcagca ggaagctcgg
     2341 cgcaaacgtt gattgtttgt ctgcgtagaa tcctctgttt gtcatatagc ttgtaatcac
     2401 gacattgttt cctttcgctt gaggtacagc gaagtgtgag taagtaaagg ttacatcgtt
     2461 aggatcaaga tccattttta acacaaggcc agttttgttc agcggcttgt atgggccagt
     2521 taaagaatta gaaacataac caagcatgta aatatcgtta gacgtaatgc cgtcaatcgt
     2581 catttttgat ccgcgggagt cagtgaacag gtaccatttg ccgttcattt taaagacgtt
     2641 cgcgcgttca atttcatctg ttactgtgtt agatgcaatc agcggtttca tcactttttt
     2701 cagtgtgtaa tcatcgttta gctcaatcat accgagagcg ccgtttgcta actcagccgt
     2761 gcgtttttta tcgctttgca gaagtttttg actttcttga cggaagaatg atgtgctttt
     2821 gccatagtat gctttgttaa ataaagattc ttcgccttgg tagccatctt cagttccagt
     2881 gtttgcttca aatactaagt atttgtggcc tttatcttct acgtagtgag gatctctcag
     2941 cgtatggttg tcgcctgagc tgtagttgcc ttcatcgatg aactgctgta cattttgata
     3001 cgtttttccg tcaccgtcaa agattgattt ataatcctct acaccgttga tgttcaaaga
     3061 gctgtctgat gctgatacgt taacttgtgc agttgtcagt gtttgtttgc cgtaatgttt
     3121 accggagaaa tcagtgtaga ataaacggat ttttccgtca gatgtaaatg tggctgaacc
     3181 tgaccattct tgtgtttggt cttttaggat agaatcattt gcatcgaatt tgtcgctgtc
     3241 tttaaagacg cggccagcgt ttttccagct gtcaatagaa gtttcgccga ctttttgata
     3301 gaacatgtaa atcgatgtgt catccgcatt tttaggatct ccggctaatg caaagacgat
     3361 gtggtagccg tgatagtttg cgacagtgcc gtcagcgttt tgtaatggcc agctgtccca
     3421 aacgtccagg ccttttgcag aagagatatt tttaattgtg gacgaatcaa attcagaaac
     3481 ttgatatttt tcattttttt gctgttcagg gatttgcagc atatcatggc gtgtaatatg
     3541 ggaaatgccg tatgtttcct tatatggctt ttggttcgtt tctttcgcaa acgcttgagt
     3601 tgcgcctcct gccagcagtg cggtagtaaa ggttaatact gttgcttgtt ttgcaaactt
     3661 tttgatgttc atcgtcgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3721 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     3781 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     3841 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     3901 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atccgcgcct tgagcctggc
     3961 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4021 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4081 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4141 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4201 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4261 ggccagccac gatagccgcg ctgcctcgtc ttggagttca ttcagggcac cggacaggtc
     4321 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     4381 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     4441 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtttgca
//
