LOCUS       sequence-424020-        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      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     601..617
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     601..617
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2145..2162)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2145..2161)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2243..3037
                     /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(2297..2316)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2907..2926
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3044..3130,3131..4465)
                     /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 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgcctcaca
      601 caggaaacag ctatgacatg attacgaatt cccttgccga cgacaccagc aacggccagg
      661 gtgatttcgc gcaggaagcg aagacggttc tgcagcgagc cgccatattc atcatcacgc
      721 tggttggtat gggccgaaat gaactggttt accaggtacc cgttcgcgca gtgaagctcc
      781 accccgtcaa aacctgcgtc caaggcattt cgggctgctt gggcgtagag cttaaccagt
      841 tcctttacct ccttggtgga cagggcacgt ggggtcgatg ggtcggccag tgcacccgtg
      901 ccggggccag tttcgatgaa caccttgacg ttgtccgcgc ggatggctga tggcgcgacg
      961 ggcgcttcac caccgggctg cagcgatgtg tgcgacacgc ggcctacatg ccaaagttga
     1021 gcaaagatca cccccccttc ggcgtgaaca gcgtcggtga ccttacgcca gccttgtatc
     1081 tgttctgggc tatgaatgcc gggtgtccag gcgtaaccct gtccccgggg ttcgatctgg
     1141 gtgccctcgg tcaccatgaa gccagcaccg ctgcgctgac ggtaataggt ggccatgagg
     1201 tcgttcgcaa tgttgccggg ttgggagctg cgcgaacgcg tcagtggcgg cagtacgata
     1261 cggttcttga gggtgtatgg gcccagctga gtggttttga aaagttctga ttgtgtcatt
     1321 tcatttacct gcatatctaa aatattggat gttgtaagct aaaaaacatg gatcaacgag
     1381 ggatatcagg gtggtttacg aatccaggct attcgtaata ttcagatatg tcaatatgta
     1441 gctttgtatt ccggcactta tgcatatctc cgcacccttg cagctcactt ctggatcagc
     1501 cgggaacgcg ccgtgcttcg cgccagggtt gcctatgtag tccagccccc cttgatctat
     1561 tcatcaagga ggctggctgc atcaggtatt gatgtttgcg ttgatgtgca gatgaagtta
     1621 gaacggcgcg tcaatatcga tgacgctaat cagcttgtga ttgacgaatt ccttgatgcc
     1681 caagccgatt agctcgcgcc cgtagccgga gcgtttgata ccaccgaagg gtaggttggc
     1741 cctcgaggtg ctgggatggt tgacgaacac cataccagtg gagatgcgtt tggccacctc
     1801 ggcgccatgc ttagtatcgg cggtgaatac cgagccgccc agaccgaacg gcgaatcgtt
     1861 ggcaatacgc accgcgtcct gctcgtccct ggcacggaag atcatcgaca ctgggccgaa
     1921 gaactcccag taataggcgg gattatcggg ctctacgtca gtcaggatag ttggttgcac
     1981 gaaggccccc agtctaggga ctctggggcc gacttcggtg gctctcgcgc cgtgagcgac
     2041 tgccacctgg atcttctgac gaatctcgtc ggcggcggcc tgcgatgaca aaggggccag
     2101 cgtggtttcc tgggcaaagg gatcaccagc cttaaaagct tggcactggc cgtcgtttta
     2161 caacgtcgtg accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca
     2221 aacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2281 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2341 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2401 gtccggtgcc ctgaatgaac tccaagacga ggcagcgcgg ctatcgtggc tggccacgac
     2461 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2521 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2581 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2641 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     2701 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2761 gctcaaggcg cggatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2821 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2881 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     2941 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3001 catcgccttc tatcgccttc ttgacgagtt cttctgagcg acgatgaaca tcaaaaagtt
     3061 tgcaaaacaa gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca
     3121 agcgtttgcg aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat
     3181 tacacgccat gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagtttc
     3241 tgaatttgat tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg acgtttggga
     3301 cagctggcca ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt
     3361 ctttgcatta gccggagatc ctaaaaatgc ggatgacaca tcgatttaca tgttctatca
     3421 aaaagtcggc gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag
     3481 cgacaaattc gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc
     3541 agccacattt acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca
     3601 ttacggcaaa caaacactga caactgcaca agttaacgta tcagcatcag acagctcttt
     3661 gaacatcaac ggtgtagagg attataaatc aatctttgac ggtgacggaa aaacgtatca
     3721 aaatgtacag cagttcatcg atgaaggcaa ctacagctca ggcgacaacc atacgctgag
     3781 agatcctcac tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg
     3841 aactgaagat ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag
     3901 cacatcattc ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc
     3961 tgagttagca aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa
     4021 agtgatgaaa ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt
     4081 ctttaaaatg aacggcaaat ggtacctgtt cactgactcc cgcggatcaa aaatgacgat
     4141 tgacggcatt acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg
     4201 cccatacaag ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga
     4261 tgtaaccttt acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat
     4321 tacaagctat atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgccgag
     4381 cttcctgctg aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca
     4441 aggacaatta acagttaaca aataatcaga ccccgtagaa aagatcaaag gatcttc
//
