LOCUS       sequence-339052-        5958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  AUTHORS   Lee SK, Keasling JD
  TITLE     A propionate-inducible expression system for enteric bacteria.
  JOURNAL   Appl Environ Microbiol. 2005 Nov . 71(11):6856-62.
  PUBMED    16269719
REFERENCE   2  (bases 1 to 5958)
  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: "Appl 
            Environ Microbiol. 2005 Nov . 71(11):6856-62."
COMMENT     Sequence Label: pPro18-km
FEATURES             Location/Qualifiers
     source          1..5958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..87)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     240..257
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     misc_feature    290..346
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2258..2277)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2415..2437)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2523..2663
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2578..2597
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2678..2695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2849..3437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2929..2948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(3548..4003)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3685..3706)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3897..3916
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(4485..5300)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4485..5300)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5208..5227
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5292..5311
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5492..5511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5730..5821)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(5840..5867)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 tttcgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcatggggag
      121 accccacact accatcggcg ctacggcgtt tcacttctga gttcggcatg gggtcaggtg
      181 ggaccaccgc gctactgccg ccaggcaaat tctgttttat cagaccgctt ctgcgttctg
      241 atttaatctg tatcaggctg aaaatcttct ctcatccgcc aaaacagcca agcttgcatg
      301 cctgcaggtc gactctagag gatccccggg taccgagctc gaattcgcta gcttgttatc
      361 aacttgttat ttgcgttgat aaagacaaag caaggggtgt gccacaattg ccttaataaa
      421 attaaatgtt taatttcaat ttgttagaga taaagtcatg aactaatccg caaatatgcg
      481 tttcagttaa cgtttcaggc aatgtttcac cgcgtttcat tgcaacaatt atgaaacaag
      541 actaaaccca atattcggtt tcttaacttt gcggtgcgct atggcacatc caccacggct
      601 taatgacgac aaaccggtta tctggacggt atctgtaacg cgcctgttcg agctgtttcg
      661 cgatatcagc ctcgagtttg atcacctggc gaacattacc cctatccagc ttggctttga
      721 aaaagcggtg acctacatcc acaagaaact ggcaaacgaa cgctgtgacg ccatcatcgc
      781 cgctggatct aacggcgcgt acctgaaaag ccgcctgtca gtgccagtta ttttgattaa
      841 accgagcggc tacgatgtgt tacaggcact ggcaaaagcc ggaaaactca cctcttctat
      901 cggcgttgtc acttatcagg aaactattcc ggcactggtg gcgtttcaaa aaacctttaa
      961 tttgcgcctc gatcaacgta gctacattac cgaagaagac gcacgcgggc agattaacga
     1021 gctaaaagct aacggcaccg aagcggtggt cggcgcgggg ctgattaccg atctggcaga
     1081 agaagccgga atgaccggaa tttttatcta ttccgccgcc accgtgcgcc aggcgttcag
     1141 cgatgcgctg gatatgacgc gcatgtcgtt acgccataac actcacgatg ccacccgcaa
     1201 cgccctgaga actcgttacg tgctgggcga tatgctcggt caatcaccac agatggaaca
     1261 agtacggcag actattttgc tgtatgcccg ctccagtgca gcggtgttga ttgaggggga
     1321 aacggggacg ggcaaagagc tggcggccca ggcgattcat cgggaatatt ttgcccgcca
     1381 cgatgcgcga cagggcaaaa agtcgcatcc gtttgttgca gtcaactgcg gggcgattgc
     1441 cgaatcgctg ctggaagcag aactgtttgg ctatgaggaa ggggcgttta ccggctcgcg
     1501 acgcggcggt cgcgccgggc tgtttgaaat cgcccacgga ggtacgctgt ttctcgatga
     1561 gattggcgaa atgccgctgc cgttgcagac ccggctgctg cgggtgctgg aagaaaaaga
     1621 ggtcacccgc gtcggcgggc atcagcctgt tccggtggat gtgcgggtca ttagcgccac
     1681 tcactgcaat ctggaagaag atatgcggca agggcagttt cgccgtgacc tgttttatcg
     1741 gctgagtatt ttgcgtctgc aattgccacc actgcgcgag cgggtggcgg atattctgcc
     1801 actggcggaa agctttttga aagtgtctct ggcggcgctc tccaccccgt tttctgccgc
     1861 attacgccag ggattacagg caagcgaaac cgtgctggtg cactacgact ggccgggcaa
     1921 tattcgtgaa ctgcgcaata tgatggagcg actggcgcta tttttaagtg tggaaccgac
     1981 gccggattta acgccgcaat ttttgcagct gctactgccg gaactggcgc gcgagtcggc
     2041 gaaaactccc gctccacgct tactgacacc acaacaggca ctggagaaat ttaatggcga
     2101 taaaacagca gcggcgaatt atttaggcat cagccggacg acgttctggc ggcggctgaa
     2161 aagctgaatc gatgataagc tgtcaaacat gagcagatcc tctacgccgg acgcatcgtg
     2221 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     2281 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     2341 gcaggccccg tggccggggg actgttgggc gccatctcct tctgcctcgc gcgtttcggt
     2401 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2461 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2521 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     2581 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2641 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2701 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2761 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2821 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2881 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2941 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3001 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3061 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3121 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3181 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3241 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3301 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3361 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3421 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3541 tccttttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3601 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3661 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3721 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3781 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3841 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3901 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3961 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaatcaa tctaaagtat
     4021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagggg ggggggggcg
     4201 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc
     4261 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt
     4321 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat
     4381 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc
     4441 agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     4501 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     4561 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     4621 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     4681 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     4741 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     4801 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     4861 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     4921 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     4981 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     5041 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     5101 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     5161 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     5221 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt
     5281 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     5341 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     5401 acgtggcttt cccccccccc ccactcaacc aagtcattct gagaatagtg tatgcggcga
     5461 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     5521 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5581 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5641 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5701 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5761 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5821 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct taatttgatg
     5881 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt
     5941 caaatccgct cccggcgg
//
