LOCUS       sequence-458366-        4478 bp    DNA     circular SYN 15-APR-2025
DEFINITION  L-arabinose inducible expression of heterologous genes, pBAD24-based
            backbone with KanR selective marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4478)
  AUTHORS   Schumann A, Gaballa A, Wiedmann M
  TITLE     Engineering of a monitorable expression system to characterize 
            beta-lactamase genes in Enterobacteriaceae.
  JOURNAL   J Microbiol Methods. 2025 Mar 18;232-234:107120. doi: 
            10.1016/j.mimet.2025.107120.
  PUBMED    40113096
REFERENCE   2  (bases 1 to 4478)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Microbiol
            Methods."; date: "2025-03-18"; volume: "232-234:107120. doi"; pages:
            " 10.1016/j.mimet.2025.107120"
COMMENT     Sequence Label: pBAD25
FEATURES             Location/Qualifiers
     source          1..4478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(281..300)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(438..460)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(626..643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(797..1385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(877..896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(1496..1951)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1633..1654)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1845..1864
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1996..2790)
                     /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(2107..2126)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2717..2736
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2791..2882)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2901..2928)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3020..3106)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3259..3276
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     promoter        complement(3390..3674)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(3448..3467)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(3701..4478,1..101)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc cgtgccggtt
       61 gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg aatcagacaa
      121 ttgacggctt gacggagtag catagggttt gcagaatccc tgcttcgtcc atttgacagg
      181 cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc cggacgcatc
      241 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
      301 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
      361 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttctgcct cgcgcgtttc
      421 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
      481 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
      541 cggggcgcag ccatgaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      601 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      661 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      721 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      781 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      841 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      901 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      961 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1021 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1081 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1141 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1201 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1261 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1321 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1381 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1441 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1501 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1561 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagataggg
     1621 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1681 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca
     1741 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1801 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1861 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1921 gccgcgctta atgcgccgct acagggcgcg taaatcaatc taaagtatat atgagtaaac
     1981 ttggtctgag agctctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     2041 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     2101 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     2161 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     2221 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
     2281 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     2341 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     2401 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     2461 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     2521 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     2581 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
     2641 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     2701 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     2761 acctgcgtgc aatccatctt gttcaatcat atgcttcctt tttcaatatt attgaagcat
     2821 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2881 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
     2941 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
     3001 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     3061 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
     3121 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
     3181 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
     3241 agaccgcttc tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca
     3301 aaacagccaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt accatggtga
     3361 attcctcctg ctagcccaaa aaaacgggta tggagaaaca gtagagagtt gcgataaaaa
     3421 gcgtcaggta ggatccgcta atcttatgga taaaaatgct atggcatagc aaagtgtgac
     3481 gccgtgcaaa taatcaatgt ggacttttct gccgtgatta tagacacttt tgttacgcgt
     3541 ttttgtcatg gctttggtcc cgctttgtta cagaatgctt ttaataagcg gggttaccgg
     3601 tttggttagc gagaagagcc agtaaaagac gcagtgacgg caatgtctga tgcaatatgg
     3661 acaattggtt tcttctctga atggcgggag tatgaaaagt atggctgaag cgcaaaatga
     3721 tcccctgctg ccgggatact cgtttaatgc ccatctggtg gcgggtttaa cgccgattga
     3781 ggccaacggt tatctcgatt tttttatcga ccgaccgctg ggaatgaaag gttatattct
     3841 caatctcacc attcgcggtc agggggtggt gaaaaatcag ggacgagaat ttgtttgccg
     3901 accgggtgat attttgctgt tcccgccagg agagattcat cactacggtc gtcatccgga
     3961 ggctcgcgaa tggtatcacc agtgggttta ctttcgtccg cgcgcctact ggcatgaatg
     4021 gcttaactgg ccgtcaatat ttgccaatac ggggttcttt cgcccggatg aagcgcacca
     4081 gccgcatttc agcgacctgt ttgggcaaat cattaacgcc gggcaagggg aagggcgcta
     4141 ttcggagctg ctggcgataa atctgcttga gcaattgtta ctgcggcgca tggaagcgat
     4201 taacgagtcg ctccatccac cgatggataa tcgggtacgc gaggcttgtc agtacatcag
     4261 cgatcacctg gcagacagca attttgatat cgccagcgtc gcacagcatg tttgcttgtc
     4321 gccgtcgcgt ctgtcacatc ttttccgcca gcagttaggg attagcgtct taagctggcg
     4381 cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc accacccgga tgcctatcgc
     4441 caccgtcggt cgcaatgttg gttttgacga tcaactct
//
