LOCUS       sequence-431356-        4497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of aldA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4497)
  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 4497)
  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: pIAR009
FEATURES             Location/Qualifiers
     source          1..4497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      977..1565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1466..1485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1577..1593
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1577..1593
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3121..3138)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3121..3137)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3219..4013
                     /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(3273..3292)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3883..3902
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4020..4106,4107..4497,1..944)
                     /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 cacaagaatg gtcaggttca gccacattta catctgacgg aaaaatccgt ttattctaca
       61 ctgatttctc cggtaaacat tacggcaaac aaacactgac aactgcacaa gttaacgtat
      121 cagcatcaga cagctctttg aacatcaacg gtgtagagga ttataaatca atctttgacg
      181 gtgacggaaa aacgtatcaa aatgtacagc agttcatcga tgaaggcaac tacagctcag
      241 gcgacaacca tacgctgaga gatcctcact acgtagaaga taaaggccac aaatacttag
      301 tatttgaagc aaacactgga actgaagatg gctaccaagg cgaagaatct ttatttaaca
      361 aagcatacta tggcaaaagc acatcattct tccgtcaaga aagtcaaaaa cttctgcaaa
      421 gcgataaaaa acgcacggct gagttagcaa acggcgctct cggtatgatt gagctaaacg
      481 atgattacac actgaaaaaa gtgatgaaac cgctgattgc atctaacaca gtaacagatg
      541 aaattgaacg cgcgaacgtc tttaaaatga acggcaaatg gtacctgttc actgactccc
      601 gcggatcaaa aatgacgatt gacggcatta cgtctaacga tatttacatg cttggttatg
      661 tttctaattc tttaactggc ccatacaagc cgctgaacaa aactggcctt gtgttaaaaa
      721 tggatcttga tcctaacgat gtaaccttta cttactcaca cttcgctgta cctcaagcga
      781 aaggaaacaa tgtcgtgatt acaagctata tgacaaacag aggattctac gcagacaaac
      841 aatcaacgtt tgcgccgagc ttcctgctga acatcaaagg caagaaaaca tctgttgtca
      901 aagacagcat ccttgaacaa ggacaattaa cagttaacaa ataatcagac cccgtagaaa
      961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1561 ggaaaaacgc ctcacacagg aaacagctat gacatgatta cgaattccct tgccgacgac
     1621 accagcaacg gccagggtga tttcgcgcag gaagcgaaga cggttctgca gcgagccgcc
     1681 atattcatca tcacgctggt tggtatgggc cgaaatgaac tggtttacca ggtacccgtt
     1741 cgcgcagtga agctccaccc cgtcaaaacc tgcgtccaag gcatttcggg ctgcttgggc
     1801 gtagagctta accagttcct ttacctcctt ggtggacagg gcacgtgggg tcgatgggtc
     1861 ggccagtgca cccgtgccgg ggccagtttc gatgaacacc ttgacgttgt ccgcgcggat
     1921 ggctgatggc gcgacgggcg cttcaccacc gggctgcagc gatgtgtgcg acacgcggcc
     1981 tacatgccaa agttgagcaa agatcacccc cccttcggcg tgaacagcgt cggtgacctt
     2041 acgccagcct tgtatctgtt ctgggctatg aatgccgggt gtccaggcgt aaccctgtcc
     2101 ccggggttcg atctgggtgc cctcggtcac catgaagcca gcaccgctgc gctgacggta
     2161 ataggtggcc atgaggtcgt tcgcaatgtt gccgggttgg gagctgcgcg aacgcgtcag
     2221 tggcggcagt acgatacggt tcttgagggt gtatgggccc agctgagtgg ttttgaaaag
     2281 ttctgattgt gtcatttcat ttacctgcat atctaaaata ttggatgttg taagctaaaa
     2341 aacatggatc aacgagggat atcagggtgg tttacgaatc caggctattc gtaatattca
     2401 gatatgtcaa tatgtagctt tgtattccgg cacttatgca tatctccgca cccttgcagc
     2461 tcacttctgg atcagccggg aacgcgccgt gcttcgcgcc agggttgcct atgtagtcca
     2521 gccccccttg atctattcat caaggaggct ggctgcatca ggtattgatg tttgcgttga
     2581 tgtgcagatg aagttagaac ggcgcgtcaa tatcgatgac gctaatcagc ttgtgattga
     2641 cgaattcctt gatgcccaag ccgattagct cgcgcccgta gccggagcgt ttgataccac
     2701 cgaagggtag gttggccctc gaggtgctgg gatggttgac gaacaccata ccagtggaga
     2761 tgcgtttggc cacctcggcg ccatgcttag tatcggcggt gaataccgag ccgcccagac
     2821 cgaacggcga atcgttggca atacgcaccg cgtcctgctc gtccctggca cggaagatca
     2881 tcgacactgg gccgaagaac tcccagtaat aggcgggatt atcgggctct acgtcagtca
     2941 ggatagttgg ttgcacgaag gcccccagtc tagggactct ggggccgact tcggtggctc
     3001 tcgcgccgtg agcgactgcc acctggatct tctgacgaat ctcgtcggcg gcggcctgcg
     3061 atgacaaagg ggccagcgtg gtttcctggg caaagggatc accagcctta aaagcttggc
     3121 actggccgtc gttttacaac gtcgtgaccg gaattgccag ctggggcgcc ctctggtaag
     3181 gttgggaagc cctgcaaaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3241 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3301 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3361 ttgtcaagac cgacctgtcc ggtgccctga atgaactcca agacgaggca gcgcggctat
     3421 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3481 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3541 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3601 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3661 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3721 ccgaactgtt cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc
     3781 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3841 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3901 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3961 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgacga
     4021 tgaacatcaa aaagtttgca aaacaagcaa cagtattaac ctttactacc gcactgctgg
     4081 caggaggcgc aactcaagcg tttgcgaaag aaacgaacca aaagccatat aaggaaacat
     4141 acggcatttc ccatattaca cgccatgata tgctgcaaat ccctgaacag caaaaaaatg
     4201 aaaaatatca agtttctgaa tttgattcgt ccacaattaa aaatatctct tctgcaaaag
     4261 gcctggacgt ttgggacagc tggccattac aaaacgctga cggcactgtc gcaaactatc
     4321 acggctacca catcgtcttt gcattagccg gagatcctaa aaatgcggat gacacatcga
     4381 tttacatgtt ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa aacgctggcc
     4441 gcgtctttaa agacagcgac aaattcgatg caaatgattc tatcctaaaa gaccaaa
//
