LOCUS       sequence-431366-        4490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of aldB-II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4490)
  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 4490)
  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: pIAR007
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1471..1488)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1471..1487)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1569..2363
                     /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(1623..1642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2233..2252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2370..2456,2457..3791)
                     /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"
     rep_origin      3824..4412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4313..4332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4424..4440
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4424..4440
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gccttcctcg gcgccaagaa ctggatgccg atggcctaca acaaggacac agggctgttc
       61 tacgtgccgt ccaacgagtg gggcatggac atctggaacg aaggcatcgc ctataagaaa
      121 ggtgcggcgt tcctcggtgc cggcttcacc atcaagccgc tcaatgaaga ctacatcggc
      181 gtgctgcgcg ccatcgaccc ggtcagcggc aaggaagtgt ggcgccacaa gaactatgcg
      241 ccgctgtggg gcggtgtgct gaccaccaag ggcaacctgg tgttcacggg cacgccagag
      301 ggcttcctgc aggcattcaa cgccaagacc ggcgacaagg tctgggaatt ccagaccggc
      361 tcgggcgtgc tcggctcgcc cgtcacctgg gaaatggacg gcgagcaata cgtttcggta
      421 gtctccggct ggggcggcgc ggtgccgctg tggggcggcg aagtggccaa acgggtcaag
      481 gacttcaacc agggcggcat gctctggacc ttcaagttgc ccaagcagtt gcagcaaacg
      541 gcaagcgtca agccataacg cctgcctact gccgcttgct gccgctgatc ggtggcaagc
      601 ggccctacta ccaaatgacg agagccccgc tgccaacgat gcattcgccc ggcacgcggg
      661 gcttcttacc atcggtgcat cgcctgtcgt gggacaggca ccgacccgca gatatcaacc
      721 gcgtcgcggc cttcgcgggc ttgcccgctc ccacaggttc accgctggtt ccagaaccgg
      781 tgaagaacct gtgggggcgg gcgagcccgc aaaagggccg gcacatgcat ggcctacccg
      841 taccaacgca ccccgcaagc tcgttccaaa gtagcattcg gtccagtccg cccccgtgca
      901 ttaatgcttc taccggaatc ctccggcaca agaagaggac ttgccctatg tggactaaac
      961 ccgcctatac cgacctgcgc attggcttcg aagtgacgat gtatttcgcc aaccgttgat
     1021 gacggcccgg ccatcgccac ggtggccgga tcctcctgga gtatgcgcct gtgaacctga
     1081 ttgatcgcca gcaggccctt gccttggggc gaggcttgcg cctggactgg gagccacgga
     1141 aggcttgcca cgtgctgttg tacgccgggg gcatcatcga gctcaatgcc agtgccggct
     1201 gggtcctcga gctgctggac ggccacagta ccgtggcaac ggtcatcgac cgcctggcac
     1261 aacgcttccc caatgtaccg gggctcgaag aggacgtact ggcgtttctg gaggtggccc
     1321 gcgccaaatc ctggatagaa tgaccggtag ccccgctttg gtctggttgc tttcgtggtg
     1381 ggatgcttta cgctggcggt tatcttccag aacaataaga acaggcttac cgatgagcca
     1441 gagtttcagc ccccttcgca aaagcttggc actggccgtc gttttacaac gtcgtgaccg
     1501 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaaca ggatgaggat
     1561 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     1621 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     1681 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     1741 atgaactcca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     1801 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     1861 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     1921 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     1981 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2041 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgga
     2101 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     2161 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     2221 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     2281 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     2341 gccttcttga cgagttcttc tgagcgacga tgaacatcaa aaagtttgca aaacaagcaa
     2401 cagtattaac ctttactacc gcactgctgg caggaggcgc aactcaagcg tttgcgaaag
     2461 aaacgaacca aaagccatat aaggaaacat acggcatttc ccatattaca cgccatgata
     2521 tgctgcaaat ccctgaacag caaaaaaatg aaaaatatca agtttctgaa tttgattcgt
     2581 ccacaattaa aaatatctct tctgcaaaag gcctggacgt ttgggacagc tggccattac
     2641 aaaacgctga cggcactgtc gcaaactatc acggctacca catcgtcttt gcattagccg
     2701 gagatcctaa aaatgcggat gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa
     2761 cttctattga cagctggaaa aacgctggcc gcgtctttaa agacagcgac aaattcgatg
     2821 caaatgattc tatcctaaaa gaccaaacac aagaatggtc aggttcagcc acatttacat
     2881 ctgacggaaa aatccgttta ttctacactg atttctccgg taaacattac ggcaaacaaa
     2941 cactgacaac tgcacaagtt aacgtatcag catcagacag ctctttgaac atcaacggtg
     3001 tagaggatta taaatcaatc tttgacggtg acggaaaaac gtatcaaaat gtacagcagt
     3061 tcatcgatga aggcaactac agctcaggcg acaaccatac gctgagagat cctcactacg
     3121 tagaagataa aggccacaaa tacttagtat ttgaagcaaa cactggaact gaagatggct
     3181 accaaggcga agaatcttta tttaacaaag catactatgg caaaagcaca tcattcttcc
     3241 gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg
     3301 gcgctctcgg tatgattgag ctaaacgatg attacacact gaaaaaagtg atgaaaccgc
     3361 tgattgcatc taacacagta acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg
     3421 gcaaatggta cctgttcact gactcccgcg gatcaaaaat gacgattgac ggcattacgt
     3481 ctaacgatat ttacatgctt ggttatgttt ctaattcttt aactggccca tacaagccgc
     3541 tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc taacgatgta acctttactt
     3601 actcacactt cgctgtacct caagcgaaag gaaacaatgt cgtgattaca agctatatga
     3661 caaacagagg attctacgca gacaaacaat caacgtttgc gccgagcttc ctgctgaaca
     3721 tcaaaggcaa gaaaacatct gttgtcaaag acagcatcct tgaacaagga caattaacag
     3781 ttaacaaata atcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3961 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4201 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4381 gatgctcgtc aggggggcgg agcctatgga aaaacgcctc acacaggaaa cagctatgac
     4441 atgattacga attcagcgag gccaagggca gctcggtgtt cgtcgcgccg
//
