LOCUS       sequence-413170-        6273 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 6273)
  AUTHORS   Edwards RA, Keller LH, Schifferli DM
  TITLE     Improved allelic exchange vectors and their use to analyze 987P 
            fimbria gene expression.
  JOURNAL   Gene. 1998 Jan 30;207(2):149-57.
  PUBMED    9511756
REFERENCE   2  (bases 1 to 6273)
  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: "Gene."; 
            date: "1998-01-30"; volume: "207(2)"; pages: "149-57"
COMMENT     Sequence Label: pRE107
FEATURES             Location/Qualifiers
     source          1..6273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(618..989)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1022..1131
                     /note="incP origin of transfer"
     primer_bind     complement(1812..1831)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1970..1992)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2133..2152
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(2209..3000,3001..3069))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2832..2851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3070..3174)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(3990..5324,5325..5411))
                     /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
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(5412..5857)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      5885..6273
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctctacgccg gacgcatcgt ggccggatcc agccgaccag gctttccacg cccgcgtgcc
       61 gctccatgtc gttcgcgcgg ttctcggaaa cgcgctgccg cgtttcgtga ttgtcacgct
      121 caagcccgta gtcccgttcg agcgtcgcgc agaggtcagc gagggcgcgg taggcccgat
      181 acggctcatg gatggtgttt cgggtcgggt gaatcttgtt gatggcgata tggatgtgca
      241 ggttgtcggt gtcgtgatgc acggcactga cgcgctgatg ctcggcgaag ccaagcccag
      301 cgcagatgcg gtcctcaatc gcgcgcaacg tctccgcgtc gggcttctct cccgcgcgga
      361 agctaaccag caggtgatag gtcttgtcgg cctcggaacg ggtgttgccg tgctgggtcg
      421 ccatcacctc ggccatgaca gcgggcaggg tgtttgcctc gcagttcgtg acgcgcacgt
      481 gacccaggcg ctcggtcttg ccttgctcgt cggtgatgta cttcaccagc tccgcgaagt
      541 cgctcttctt gatggagcgc atggggacgt gcttggcaat cacgcgcacc ccccggccgt
      601 tttagcggct aaaaaagtca tggctctgcc ctcgggcgga ccacgcccat catgaccttg
      661 ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg cgaggatcgt ggcatcaccg
      721 aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc
      781 aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt
      841 ttgtagccct ggccgacggc cagcaggtag gccgacaggc tcatgccggc cgccgccgcc
      901 ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc
      961 ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta
     1021 gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca
     1081 gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc
     1141 cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg
     1201 aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg
     1261 gaaaagcgct gcttccctgc tgttttgtgg aatatctacc gactggaaac aggcaaatgc
     1321 aggaaattac tgaactgagg ggacaggcga gagacgatgc caaagagcta caccgacgag
     1381 ctggccgagt gggttgaatc ccgcgcggcc aagaagcgcc ggcgtgatga ggctgcggtt
     1441 gcgttcctgg cggtgagggc ggatgtcgag gcggcgttag cgtccggcta tgcgctcgtc
     1501 accatttggg agcacatgcg ggaaacgggg aaggtcaagt tctcctacga gacgttccgc
     1561 tcgcacgcca ggcggcacat caaggccaag cccgccgatg tgcccgcacc gcaggccaag
     1621 gctgcggaac ccgcgccggc acccaagacg ccggagccac ggcggccgaa gcaggggggc
     1681 aaggctgaaa agccggcccc cgctgcggcc ccgaccggct tcaccttcaa cccaacaccg
     1741 gacaaaaagg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     1801 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     1861 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     1921 gggcgccatc tccttgctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1981 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2041 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     2101 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     2161 gcaccataaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2521 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2821 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3181 tccccgaaaa gtgccacctg cagatctgca ggtcgacgga tcccaagctt cttctagagg
     3241 taccgcatgc gatatcgagc tctcccggga attcatgcag ttcacttaca ccgcttctca
     3301 acccggtacg caccagaaaa tcattgatat ggccatgaat ggcgttggat gccgggcaac
     3361 agcccgcatt atgggcgttg gcctcaacac gattttacgt cacttaaaaa actcaggccg
     3421 cagtcggtaa cctcgcgcat acagccgggc agtgacgtca tcgtctgcgc ggaaatggac
     3481 gaacagtggg gctatgtcgg ggctaaatcg cgccagcgct ggctgtttta cgcgtatgac
     3541 agtctccgga agacggttgt tgcgcacgta ttcggtgaac gcactatggc gacgctgggg
     3601 cgtcttatga gcctgctgtc accctttgac gtggtgatat ggatgacgga tggctggccg
     3661 ctgtatgaat cccgcctgaa gggaaagctg cacgtaatca gcaagcgata tacgcagcga
     3721 attgagcggc ataacctgaa tctgaggcag cacctggcac ggctgggacg gaagtcgctg
     3781 tcgttctcaa aatcggtgga gctgcatgac aaagtcatcg ggcattatct gaacataaaa
     3841 cactatcaat aagttggagt cattaccaaa aggttaggaa tacggttagc catttgcctg
     3901 cttttatata gttcatatgg gattcacctt tatgttgata agaaataaaa gaaaatgcca
     3961 ataggatatc ggcattttct tttgcgtttt tatttgttaa ctgttaattg tccttgttca
     4021 aggatgctgt ctttgacaac agatgttttc ttgcctttga tgttcagcag gaagctgggc
     4081 gcaaacgttg attgtttgtc tgcgtagaat cctctgtttg tcatatagct tgtaatcacg
     4141 acattgtttc ctttcgcttg aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta
     4201 ggatcaagat ccatttttaa cacaaggcca gttttgttca gcggcttgta tgggccagtt
     4261 aaagaattag aaacataacc aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc
     4321 atttttgatc cgcgggagtc agtgaacaga taccatttgc cgttcatttt aaagacgttc
     4381 gcgcgttcaa tttcatctgt tactgtgtta gatgcaatca gcggtttcat cacttttttc
     4441 agtgtgtaat catcgtttag ctcaatcata ccgagagcgc cgtttgctaa ctcagccgtg
     4501 cgttttttat cgctttgcag aagtttttga ctttcttgac ggaagaatga tgtgcttttg
     4561 ccatagtatg ctttgttaaa taaagattct tcgccttggt agccatcttc agttccagtg
     4621 tttgcttcaa atactaagta tttgtggcct ttatcttcta cgtagtgagg atctctcagc
     4681 gtatggttgt cgcctgagct gtagttgcct tcatcgatga actgctgtac attttgatac
     4741 gtttttccgt caccgtcaaa gattgattta taatcctcta caccgttgat gttcaaagag
     4801 ctgtctgatg ctgatacgtt aacttgtgca gttgtcagtg tttgtttgcc gtaatgttta
     4861 ccggagaaat cagtgtagaa taaacggatt tttccgtcag atgtaaatgt ggctgaacct
     4921 gaccattctt gtgtttggtc ttttaggata gaatcatttg catcgaattt gtcgctgtct
     4981 ttaaagacgc ggccagcgtt tttccagctg tcaatagaag tttcgccgac tttttgatag
     5041 aacatgtaaa tcgatgtgtc atccgcattt ttaggatctc cggctaatgc aaagacgatg
     5101 tggtagccgt gatagtttgc gacagtgccg tcagcgtttt gtaatggcca gctgtcccaa
     5161 acgtccaggc cttttgcaga agagatattt ttaattgtgg acgaatcgaa ctcaggaact
     5221 tgatattttt catttttttg ctgttcaggg atttgcagca tatcatggcg tgtaatatgg
     5281 gaaatgccgt atgtttcctt atatggcttt tggttcgttt ctttcgcaaa cgcttgagtt
     5341 gcgcctcctg ccagcagtgc ggtagtaaag gttaatactg ttgcttgttt tgcaaacttt
     5401 ttgatgttca tcgttcatgt ctcctttttt atgtactgtg ttagcggtct gcttcttcca
     5461 gccctcctgt ttgaagatgg caagttagtt acgcacaata aaaaaagacc taaaatatgt
     5521 aaggggtgac gccaaagtat acactttgcc ctttacacat tttaggtctt gcctgcttta
     5581 tcagtaacaa acccgcgcga tttacttttc gacctcattc tattagactc tcgtttggat
     5641 tgcaactggt ctattttcct cttttgtttg atagaaaatc ataaaaggat ttgcagacta
     5701 cgggcctaaa gaactaaaaa atctatctgt ttcttttcat tctctgtatt ttttatagtt
     5761 tctgttgcat gggcataaag ttgccttttt aatcacaatt cagaaaatat cataatatct
     5821 catttcacta aataatagtg aacggcaggt atatgtgatg ggttaaaaag gatcagaatt
     5881 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
     5941 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
     6001 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
     6061 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
     6121 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
     6181 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
     6241 ctatcaacag gttgaactgc tgatcttcag atc
//
