LOCUS       sequence-431369-        4537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of aldB-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4537)
  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 4537)
  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: pIAR002
FEATURES             Location/Qualifiers
     source          1..4537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..103
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(221..307,308..1642)
                     /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      1675..2263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2164..2183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2275..2291
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2275..2291
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3859..3876)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3859..3875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3957..4537,1..214)
                     /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(4011..4030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
       61 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
      121 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
      181 cgccttctat cgccttcttg acgagttctt ctgagcgacg atgaacatca aaaagtttgc
      241 aaaacaagca acagtattaa cctttactac cgcactgctg gcaggaggcg caactcaagc
      301 gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca tacggcattt cccatattac
      361 acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc aagtttctga
      421 atttgattcg tccacaatta aaaatatctc ttctgcaaaa ggcctggacg tttgggacag
      481 ctggccatta caaaacgctg acggcactgt cgcaaactat cacggctacc acatcgtctt
      541 tgcattagcc ggagatccta aaaatgcgga tgacacatcg atttacatgt tctatcaaaa
      601 agtcggcgaa acttctattg acagctggaa aaacgctggc cgcgtcttta aagacagcga
      661 caaattcgat gcaaatgatt ctatcctaaa agaccaaaca caagaatggt caggttcagc
      721 cacatttaca tctgacggaa aaatccgttt attctacact gatttctccg gtaaacatta
      781 cggcaaacaa acactgacaa ctgcacaagt taacgtatca gcatcagaca gctctttgaa
      841 catcaacggt gtagaggatt ataaatcaat ctttgacggt gacggaaaaa cgtatcaaaa
      901 tgtacagcag ttcatcgatg aaggcaacta cagctcaggc gacaaccata cgctgagaga
      961 tcctcactac gtagaagata aaggccacaa atacttagta tttgaagcaa acactggaac
     1021 tgaagatggc taccaaggcg aagaatcttt atttaacaaa gcatactatg gcaaaagcac
     1081 atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac gcacggctga
     1141 gttagcaaac ggcgctctcg gtatgattga gctaaacgat gattacacac tgaaaaaagt
     1201 gatgaaaccg ctgattgcat ctaacacagt aacagatgaa attgaacgcg cgaacgtctt
     1261 taaaatgaac ggcaaatggt acctgttcac tgactcccgc ggatcaaaaa tgacgattga
     1321 cggcattacg tctaacgata tttacatgct tggttatgtt tctaattctt taactggccc
     1381 atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc ctaacgatgt
     1441 aacctttact tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg tcgtgattac
     1501 aagctatatg acaaacagag gattctacgc agacaaacaa tcaacgtttg cgccgagctt
     1561 cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc ttgaacaagg
     1621 acaattaaca gttaacaaat aatcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1681 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1741 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1801 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1861 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1921 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1981 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2041 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2101 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2161 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2221 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcct cacacaggaa
     2281 acagctatga catgattacg aattcgcgat ggcggtgatg atgaacatca gcttcagtgc
     2341 ctcacccacg ccgaaaatgt ggatgctgat gaacacgatg tagaacgcca ggtagatcat
     2401 ccagccgccg ataccgaaca gcgattggca gtaggcgccg atgaacaccg cgatggcggc
     2461 tggggcgatg gcgtattcga tcaggatcgc cgtgccggtc aaaaagccgc cccacggtcc
     2521 gaacgcgctg cgggcaaagc cgtagccgcc gccagcggtg gggatcatcg acgacagttc
     2581 ggccagcgag aagcacatgc acaggtacat ggttgccatc agcaaggtgg ccaggaacat
     2641 gccgccccag ccaccctgcg ccaggccgaa gttccagccg gcgtaatcgc cggaaatgac
     2701 gtacgccacg cccaggccga ccagcagtac ccagccggcg gcgccttttt tcagttcacg
     2761 ttgctggaaa tagtcagagc cgactttctc gaagtcgacg gaggagcctg ccggcgatcc
     2821 ggcggaatgt tcgcttggca tagattcacc tgtttttttt cttggtggcc ggaagggttc
     2881 cgggccgatg gtgatgggtg ctgcaggaag cgagccagag cggttggtgc acggttaccg
     2941 ccctgggttt tgtgtaaagc aagtccggcc tcttcgcggg cttgcccgct cccacaggta
     3001 ccgcaccggc tctggaacca acggtgaacc tgtaggagcg ggtttacccg cgaagaaggc
     3061 gacgcggtct agaagaagga tatcgtgtgt ctccttggta ttgtgctttt tagggagttg
     3121 aaaagcatag agcaagggtt gggccagtgt tggcggtccc aggcaaatca atgacttggg
     3181 tcatttttac gggcctgttc aaggaccgct gtgccggcgc cggtacgcct ggggtgacat
     3241 tttgtgccgt ttgcgacaca tcacaggcgt gtgcattgca aagcctaacc gggtggagga
     3301 tgctgggctg acgctctatc tgcggagaat tacaacaatg cagagcaatc acttcactcg
     3361 ccatgcccgg caggtccata gcgttgccca tggcggggcc ggggagggcg gcagtgatcc
     3421 gtccatcgcc cgttcctggc tgcgctgcct ggaggattac cacctcgacc cggcggtgat
     3481 cgaagcgccg gtggtgcttg agcatgggcg cctgctggaa agccgcgagc gcctgcacca
     3541 ggtgctgcaa attgccgacc atgaaatgaa cagcctgcac cagcagcttt ccggcgcagg
     3601 ccacgcggtg ctgctgaccg acgcccgcgg ggtgatcctc aattgcgtca gtgcccccgc
     3661 cgagcggcgc agcttcgagc gtgccgggct gtggctgggc gctgactgga gcgaggcgcg
     3721 cgagggcacc aatggcatcg gcacatgtct ggtcgagcgc caggcgctga ccatccatca
     3781 gaacgagcac ttccgtggcc gtcacaccgg gctgacctgc tcggccagcc cggtgttcga
     3841 cccgcatgga agcttggcac tggccgtcgt tttacaacgt cgtgaccgga attgccagct
     3901 ggggcgccct ctggtaaggt tgggaagccc tgcaaacagg atgaggatcg tttcgcatga
     3961 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4021 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4081 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactccaag
     4141 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4201 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4261 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4321 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4381 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4441 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcggatg cccgacggcg
     4501 aggatctcgt cgtgacccat ggcgatgcct gcttgcc
//
