LOCUS       sequence-233791-        5428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression vector with Gentamycin selection, based on 
            pBAD33.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5428)
  AUTHORS   Jimenez N, Lacasta A, Vilches S, Reyes M, Vazquez J, Aquillini E, 
            Merino S, Regue M, Tomas JM
  TITLE     Genetics and proteomics of Aeromonas salmonicida lipopolysaccharide 
            core biosynthesis.
  JOURNAL   J Bacteriol. 2009 Apr;191(7):2228-36. doi: 10.1128/JB.01395-08. Epub
            2009 Jan 16.
  PUBMED    19151135
REFERENCE   2  (bases 1 to 5428)
  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"; doi: 
            "10.1128/JB.01395-08"; journalName: "J Bacteriol"; date: "2009-04"; 
            volume: "191"; issue: "7"; pages: "2228-36"
COMMENT     Sequence Label: pBAD33-Gm
FEATURES             Location/Qualifiers
     source          1..5428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(83..961)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        988..1272
                     /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     1195..1214
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    1293..1349
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1382..1399)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1552..1638
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1730..1757
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1776..1867
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2325..2780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2412..2431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2622..2643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    3427..3460
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3503..4036)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4225..4230,4231..4247,4248..4253))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    4273..4306
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4557..4574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4691..5236)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4771..4790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5312..5331
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtgcctgtca aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt
       61 caattgtctg attcgttacc aattatgaca acttgacggc tacatcattc actttttctt
      121 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
      181 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
      241 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
      301 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
      361 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
      421 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
      481 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
      541 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
      601 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
      661 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
      721 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
      781 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
      841 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
      901 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
      961 tacttttcat actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt
     1021 gccgtcactg cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta
     1081 aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct
     1141 ataatcacgg cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca
     1201 tagcattttt atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta
     1261 ctgtttctcc atacccgttt ttttgggcta gcgaattcga gctcggtacc cggggatcct
     1321 ctagagtcga cctgcaggca tgcaagcttg gctgttttgg cggatgagag aagattttca
     1381 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1441 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1501 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1561 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1621 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     1681 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     1741 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     1801 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1861 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1921 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1981 gggtgcagca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2041 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2101 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2161 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2221 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2281 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
     2341 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2401 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2461 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2521 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2581 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2641 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2701 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2761 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2821 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2881 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2941 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3001 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3061 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3121 cagtcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     3181 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     3241 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     3301 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     3361 cagtgggtac cgagctcacc ggttaacacg cgtacgtagg gcccgcggta tcgataagct
     3421 ggatccataa cttcgtataa tgtatgctat acgaagttat ctacgcgtcg gccgggaagc
     3481 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     3541 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     3601 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     3661 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     3721 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     3781 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     3841 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     3901 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     3961 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     4021 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     4081 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     4141 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagctgcgt
     4201 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caattcgaga
     4261 attgacgcgt agataacttc gtataatgta tgctatacga agttatgcgg ccgccatatg
     4321 catccatggt accaattgta cagctgaaga gctcggtacc ctgtccctcc tgttcagcta
     4381 ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat
     4441 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     4501 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     4561 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     4621 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     4681 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     4741 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     4801 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     4861 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     4921 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     4981 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     5041 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     5101 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     5161 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     5221 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct aggctcatga
     5281 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5341 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat
     5401 gtttgacagc ttatcatcga tgcataat
//
