LOCUS       sequence-431361-        4526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of modABC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4526)
  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 4526)
  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: pIAR010
FEATURES             Location/Qualifiers
     source          1..4526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(332..1126)
                     /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(443..462)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1053..1072
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1207..1224
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1208..1224
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2781..2797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2781..2797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(2809..3397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2889..2908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3430..4526,1..238,239..325))
                     /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 atgcaaagac gatgtggtag ccgtgatagt ttgcgacagt gccgtcagcg ttttgtaatg
       61 gccagctgtc ccaaacgtcc aggccttttg cagaagagat atttttaatt gtggacgaat
      121 caaattcaga aacttgatat ttttcatttt tttgctgttc agggatttgc agcatatcat
      181 ggcgtgtaat atgggaaatg ccgtatgttt ccttatatgg cttttggttc gtttctttcg
      241 caaacgcttg agttgcgcct cctgccagca gtgcggtagt aaaggttaat actgttgctt
      301 gttttgcaaa ctttttgatg ttcatcgtcg ctcagaagaa ctcgtcaaga aggcgataga
      361 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
      421 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
      481 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
      541 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatccgcg
      601 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
      661 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
      721 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
      781 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
      841 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
      901 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttggagt tcattcaggg
      961 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     1021 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     1081 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     1141 atcctgtttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggtca
     1201 cgacgttgta aaacgacggc cagtgccaag cttgcagaaa ctgattgacc tcgctggagc
     1261 cgacactatg gcgctggcca tcttcatcca ggtactgaaa cagtgcctgc ccaggcagtt
     1321 ccatgcagcg tttgagtagg ttggccaagc gccggtcgtt gagggtgacg ttgtgttcga
     1381 cgccgcgctt gccgcggaac tggaagcgga tagtgctgcc tttgacctgc acatggcgat
     1441 tgcgcagggt ggtcaatccg tacgaccggt tatcgcgcag gtagcgctgg ttgccgatgc
     1501 ggatcagggt gtggtccagc aggctcacca ccagcgccat gactttttcc cggtccagcc
     1561 ctggccgcgc caagtgggct tccagctgtg tgcgcagctt tggcagggct tgggcgaacg
     1621 caagcatgcg cccgtatttg tgctggtcgc gcagttcgcg ccactgtgcg tggtagcggt
     1681 actgcttgcg gccacgggca tcgcggccgg tggcttgcag gtggccttgc ggatcggcgc
     1741 agatccacac atccgtgtag gccgggggga tcaccaatgc ggcgatgcgc gcaagggttt
     1801 cgctgtcgcg tacccgttgg ccatcggcat ccaggtagat gaaacggtcg cgccagcgcc
     1861 gacgggtcag gcccggctga ctgtcgtcaa cgtagtgcag ggtgcgcggc agggggcagt
     1921 caagcatggg cggcgggcct cgatcaggtc actgatcaat ggacaatacg cgcgctgagg
     1981 atgctcaacc gatatcgaca gctccttgag gttatcgtta tagccttaac tatatagcga
     2041 taagccattg gccgtgaaga ggccatgcct gcctgtcaga atgcctggtg accgatcact
     2101 gccaatgcct taccggcaat ctcccgggtc agctcacccg ccggcatgtg ccggcctagc
     2161 cgtaacgcct gccccgacca cagattgctg aaatcactct taccctgcgg atcggtgatc
     2221 gcccgtaaag gcatcaatgc cccgcccgcc agcgggaagc gcggtgcaag ctcgctcatc
     2281 ggccccagct cgcgcatgag gcgattgttg atgccgcgcg ccgggcggcc ggtgaacagg
     2341 ttggtcaggg cggtgtcgct ggcaggcgcg ctgtccagcg cctggcgatg cgctggcgac
     2401 accttggcct cggggcagaa caggtaagcc gtgccgatct gcactgccga ggcacccagg
     2461 gccagtgccg ccaacagccc gcggtggtcg ccgataccac cggcggcaat caccggcacg
     2521 ccgacggcat cggccacctg cggtacgagc gcgaaggtgc ctatctggct ggtgatgtca
     2581 tcgctgagga acatgccgcg atggccaccg gcttcatacc ccatggcgat gatcgcatcg
     2641 cagccattgc gctccagcca ggccgcttct tccacggtcg tggcgctgga cagtaccttg
     2701 gcaccgctgg cctttacccg ctgcagcagt tcggcctgcg gcaggccgaa atggaagctc
     2761 accaccgaat tcgtaatcat gtcatagctg tttcctgtgt gaggcgtttt tccataggct
     2821 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2881 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2941 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3001 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3061 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3121 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3181 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3241 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3301 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3421 ggggtctgat tatttgttaa ctgttaattg tccttgttca aggatgctgt ctttgacaac
     3481 agatgttttc ttgcctttga tgttcagcag gaagctcggc gcaaacgttg attgtttgtc
     3541 tgcgtagaat cctctgtttg tcatatagct tgtaatcacg acattgtttc ctttcgcttg
     3601 aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat ccatttttaa
     3661 cacaaggcca gttttgttca gcggcttgta tgggccagtt aaagaattag aaacataacc
     3721 aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc atttttgatc cgcgggagtc
     3781 agtgaacagg taccatttgc cgttcatttt aaagacgttc gcgcgttcaa tttcatctgt
     3841 tactgtgtta gatgcaatca gcggtttcat cacttttttc agtgtgtaat catcgtttag
     3901 ctcaatcata ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat cgctttgcag
     3961 aagtttttga ctttcttgac ggaagaatga tgtgcttttg ccatagtatg ctttgttaaa
     4021 taaagattct tcgccttggt agccatcttc agttccagtg tttgcttcaa atactaagta
     4081 tttgtggcct ttatcttcta cgtagtgagg atctctcagc gtatggttgt cgcctgagct
     4141 gtagttgcct tcatcgatga actgctgtac attttgatac gtttttccgt caccgtcaaa
     4201 gattgattta taatcctcta caccgttgat gttcaaagag ctgtctgatg ctgatacgtt
     4261 aacttgtgca gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat cagtgtagaa
     4321 taaacggatt tttccgtcag atgtaaatgt ggctgaacct gaccattctt gtgtttggtc
     4381 ttttaggata gaatcatttg catcgaattt gtcgctgtct ttaaagacgc ggccagcgtt
     4441 tttccagctg tcaatagaag tttcgccgac tttttgatag aacatgtaaa tcgatgtgtc
     4501 atccgcattt ttaggatctc cggcta
//
