LOCUS       sequence-431368-        4506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of sadII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  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 4506)
  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: pIAR005
FEATURES             Location/Qualifiers
     source          1..4506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(338..1132)
                     /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(449..468)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1059..1078
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1213..1230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1214..1230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2767..2783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2767..2783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(2795..3383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2875..2894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3416..4506,1..244,245..331))
                     /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 cggctaatgc aaagacgatg tggtagccgt gatagtttgc gacagtgccg tcagcgtttt
       61 gtaatggcca gctgtcccaa acgtccaggc cttttgcaga agagatattt ttaattgtgg
      121 acgaatcaaa ttcagaaact tgatattttt catttttttg ctgttcaggg atttgcagca
      181 tatcatggcg tgtaatatgg gaaatgccgt atgtttcctt atatggcttt tggttcgttt
      241 ctttcgcaaa cgcttgagtt gcgcctcctg ccagcagtgc ggtagtaaag gttaatactg
      301 ttgcttgttt tgcaaacttt ttgatgttca tcgtcgctca gaagaactcg tcaagaaggc
      361 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
      421 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
      481 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
      541 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
      601 tccgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
      661 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
      721 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
      781 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
      841 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
      901 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tggagttcat
      961 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     1021 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     1081 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     1141 atcctcatcc tgtttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc
     1201 cggtcacgac gttgtaaaac gacggccagt gccaagcttg actccatttc gtgaatcttg
     1261 cccggtaccg cctggtcctg cacgaaccag ttttcgaagt gatggcggta tgaacagttg
     1321 gcgcggaagg cgtagatgct ttcaccgttc acgtcctgcg cgcgggtcac cggtgtatgg
     1381 ttgagcggcg caatgaccat catctcttcc tcgaacaccg gtatgccctc cagcgtcggg
     1441 tgcagcaccg ggccgtcgac gaacgcggcc accaggcggc cagacaacac gccttcgagc
     1501 atggtcccgg agggcccggt ggacaggtca aggtcaacct tggggtagcg ctggttgtag
     1561 gcggccagca gcgccgggat gcgcaccgcc gccgtgcttt ccagcgatcc gagggcaaag
     1621 gtgccctgcg ggtcttcacc cgccacggtc agccgcgcct catggaccag gtcgaggatg
     1681 cgccgggtgt attcgaggaa gttccaaccc gcaggcgaca ggcgcaggcg gctcttctcg
     1741 cgaatgaaca gctctacacc caggtcctcc tccagttgct tgatgcgcgt ggtcaggttc
     1801 gacggcaccc gatggatgtg ctgcgcggcg gcgctgatgc tgccctgctc tgccacagcc
     1861 ttgaagatct ccagttgcac caggtccaca gccattctcc aaacgtgaat gtttcgctca
     1921 ttattattca gttttcatta aagccatagc ccactagtct ggcctcactg attaacaaca
     1981 tcgagagtgt cccgccgata tcttcatgac cctggggctg ctttgcggcc ccagctgctt
     2041 caatcccgaa cctggtcctt gacccagtcc agcatccccc ccggcacgat caattcatgc
     2101 cgcgcccgcg aacaggcggt atacaacccc gccagcatac gcgcccgttc attgcggtta
     2161 cccgccacct gtggcgccac catcagctcc ggcgacacca tcacccgggc aaactccatg
     2221 ttcttcacat cccgtactcg ccccaggaac agcttcggcg ccttgtccca gcggtaacgg
     2281 ctcttggctt tggcaaagtg ctccggcgtg tagcctttgc gcagcatgtt ggccacggcg
     2341 acgaacgctt tgtcatcgcc cttgtcctgt tccagcgcct gccagctggc atagcggaac
     2401 agcatcgggt ggcgcggccg ggtgccgtag cggtacagct cgatgcagtc ctcgacgaac
     2461 agttcaaagt ccttgcgagc gcttttcagc aacaagaacg gcaccccctg atgcgtgagg
     2521 cgctggaacc agccaaacag gccccattca tcatcgacga taagcgccgt gggctgttcc
     2581 ggcaccatca ccgtatcgaa gaagctgacc cgggtgtagt gctcggcgct gccactgaag
     2641 gccgcctgga tcgtggccgg gtgcgcctgg atcaacgggt tgagcacatt gtccattgcc
     2701 ggcccggcgc gcagtgaatg gtcgatgcag cgttgccgga tgaagctgcc atgaattcgt
     2761 aatcatgtca tagctgtttc ctgtgtgagg cgtttttcca taggctccgc ccccctgacg
     2821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgattatt
     3421 tgttaactgt taattgtcct tgttcaagga tgctgtcttt gacaacagat gttttcttgc
     3481 ctttgatgtt cagcaggaag ctcggcgcaa acgttgattg tttgtctgcg tagaatcctc
     3541 tgtttgtcat atagcttgta atcacgacat tgtttccttt cgcttgaggt acagcgaagt
     3601 gtgagtaagt aaaggttaca tcgttaggat caagatccat ttttaacaca aggccagttt
     3661 tgttcagcgg cttgtatggg ccagttaaag aattagaaac ataaccaagc atgtaaatat
     3721 cgttagacgt aatgccgtca atcgtcattt ttgatccgcg ggagtcagtg aacaggtacc
     3781 atttgccgtt cattttaaag acgttcgcgc gttcaatttc atctgttact gtgttagatg
     3841 caatcagcgg tttcatcact tttttcagtg tgtaatcatc gtttagctca atcataccga
     3901 gagcgccgtt tgctaactca gccgtgcgtt ttttatcgct ttgcagaagt ttttgacttt
     3961 cttgacggaa gaatgatgtg cttttgccat agtatgcttt gttaaataaa gattcttcgc
     4021 cttggtagcc atcttcagtt ccagtgtttg cttcaaatac taagtatttg tggcctttat
     4081 cttctacgta gtgaggatct ctcagcgtat ggttgtcgcc tgagctgtag ttgccttcat
     4141 cgatgaactg ctgtacattt tgatacgttt ttccgtcacc gtcaaagatt gatttataat
     4201 cctctacacc gttgatgttc aaagagctgt ctgatgctga tacgttaact tgtgcagttg
     4261 tcagtgtttg tttgccgtaa tgtttaccgg agaaatcagt gtagaataaa cggatttttc
     4321 cgtcagatgt aaatgtggct gaacctgacc attcttgtgt ttggtctttt aggatagaat
     4381 catttgcatc gaatttgtcg ctgtctttaa agacgcggcc agcgtttttc cagctgtcaa
     4441 tagaagtttc gccgactttt tgatagaaca tgtaaatcga tgtgtcatcc gcatttttag
     4501 gatctc
//
