LOCUS       sequence-403719-        5424 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 5424)
  AUTHORS   Lee CY, Buranen SL, Ye ZH
  TITLE     Construction of single-copy integration vectors for Staphylococcus 
            aureus.
  JOURNAL   Gene. 1991 Jul 15. 103(1):101-5.
  PUBMED    1652539
REFERENCE   2  (bases 1 to 5424)
  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. 1991 
            Jul 15. 103(1):101-5."
COMMENT     Sequence Label: pLI50
FEATURES             Location/Qualifiers
     source          1..5424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..56
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    71..127
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             1726..2730
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     primer_bind     complement(3236..3258)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3344..3484
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3399..3418
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3499..3516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3670..4258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3750..3769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4429..5220,5221..5289))
                     /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     5052..5071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5290..5394)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
       61 tcgtcttcaa gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
      121 caagcttctg taggttttta ggcataaaac tatatgattt acccctaaat ctttaaaatg
      181 ccccttaaaa ttcaaaataa aggcatttaa aatttaaata tttcttgtga taaagtttgt
      241 taaaaaggag tggttttatg actgttatgt ggttatcgat tataggtatg tggttttgta
      301 ttggaatggc attttttgct atcaaggtta ttaaaaataa aaattagacc acgcatttat
      361 gccgagaaaa tttattgtgc gttgagaaga acccttaact aaacttgcag acgaatgtcg
      421 gcatagcgtg agctattaag ccgaccattc gacaagtttt gggattgtta agggttccga
      481 ggctcaacgt caataaagca attggaataa agaagcgaaa aaggagaagt cggttcagaa
      541 aaagaaggat atggatctgg agctgtaata taaaaacctt cttcaactaa cggggcaggt
      601 tagtgacatt agaaaaccga ctgtaaaaag tacagtcggc attatctcat attataaaag
      661 ccagtcatta ggcctatctg acaattcctg aatagagttc ataaacaatc ctgcatgata
      721 accatcacaa acagaatgat gtacctgtaa agatagcggt aaatatattg aattaccttt
      781 attaatgaat tttcctgctg taataatggg tagaaggtaa ttactattat tattgatatt
      841 taagttaaac ccagtaaatg aagtccatgg aataatagaa agagaaaaag cattttcagg
      901 tataggtgtt ttgggaaaca atttccccga accattatat ttctctacat cagaaaggta
      961 taaatcataa aactctttga agtcattctt tacaggagtc caaataccag agaatgtttt
     1021 agatacacca tcaaaaattg tataaagtgg ctctaactta tcccaataac ctaactctcc
     1081 gtcgctattg taaccagttc taaaagctgt atttgagttt atcacccttg tcactaagaa
     1141 aataaatgca gggtaaaatt tatatccttc ttgttttatg tttcggtata aaacactaat
     1201 atcaatttct gtggttatac taaaagtcgt ttgttggttc aaataatgat taaatatctc
     1261 ttttctcttc caattgtcta aatcaatttt attaaagttc atttgatatg cctcctaaat
     1321 ttttatctaa agtgaattta ggaggcttac ttgtctgctt tcttcattag aatcaatcct
     1381 tttttaaaag tcaatattac tgtaacataa atatatattt taaaaatatc ccactttatc
     1441 caattttcgt ttgttgaact aatgggtgct ttagttgaag aataaaagac cacattaaaa
     1501 aatgtggtct tttgtgtttt tttaaaggat ttgagcgtag cgaaaaatcc ttttctttct
     1561 tatcttgata catatagaaa taacgtcatt tttattttag ttgctgaaag gtgcgttgaa
     1621 gtgttggtat gtatgtgttt taaagtattg aaaaccctta aaattggttg cacagaaaaa
     1681 ccccatctgt taaagttata agtgactaaa caaataacta aatagatggg ggtttctttt
     1741 aatattatgt gtcctaatag tagcatttat tcagatgaaa aatcaagggt tttagtggac
     1801 aagacaaaaa gtggaaaagt gagaccatgg agagaaaaga aaatcgctaa tgttgattac
     1861 tttgaacttc tgcatattct tgaatttaaa aaggctgaaa gagtaaaaga ttgtgctgaa
     1921 atattagagt ataaacaaaa tcgtgaaaca ggcgaaagaa agttgtatcg agtgtggttt
     1981 tgtaaatcca ggctttgtcc aatgtgcaac tggaggagag caatgaaaca tggcattcag
     2041 tcacaaaagg ttgttgctga agttattaaa caaaagccaa cagttcgttg gttgtttctc
     2101 acattaacag ttaaaaatgt ttatgatggc gaagaattaa ataagagttt gtcagatatg
     2161 gctcaaggat ttcgccgaat gatgcaatat aaaaaaatta ataaaaatct tgttggtttt
     2221 atgcgtgcaa cggaagtgac aataaataat aaagataatt cttataatca gcacatgcat
     2281 gtattggtat gtgtggaacc aacttatttt aagaatacag aaaactacgt gaatcaaaaa
     2341 caatggattc aattttggaa aaaggcaatg aaattagact atgatccaaa tgtaaaagtt
     2401 caaatgattc gaccgaaaaa taaatataaa tcggatatac aatcggcaat tgacgaaact
     2461 gcaaaatatc ctgtaaagga tacggatttt atgaccgatg atgaagaaaa gaatttgaaa
     2521 cgtttgtctg atttggagga aggtttacac cgtaaaaggt taatctccta tggtggtttg
     2581 ttaaaagaaa tacataaaaa attaaacctt gatgacacag aagaaggcga tttgattcat
     2641 acagatgatg acgaaaaagc cgatgaagat ggattttcta ttattgcaat gtggaattgg
     2701 gaacggaaaa attattttat taaagagtag ttcaacaaac gggccagttt gttgaagatt
     2761 agatgctata attgttatta aaaggattga aggatgctta ggaagacgag ttattaatag
     2821 ctgaataaga acggtgctct ccaaatattc ttatttagaa aagcaaatct aaaattatct
     2881 gaaaagggaa tgagaatagt gaatggacca ataataatga ctagagaaga aagaatgaag
     2941 attgttcatg aaattaagga acgaatattg gataaatatg gggatgatgt taaggctatt
     3001 ggtgtttatg gctctcttgg tcgtcagact gatgggccct attcggatat tgagatgatg
     3061 tgtgtcatgt caacagagga agcagagttc agccatgaat ggacaaccgg aaagagcgaa
     3121 aatgcctcac atttgtgcca cctaaaaagg agcgatttac atatgagtta tgcagtttgt
     3181 agaatgcaaa aagtgaaatc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     3241 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     3301 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     3361 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     3421 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     3481 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4081 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4321 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4501 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4561 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4621 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4681 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4741 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4801 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4861 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4921 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4981 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5041 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5101 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5161 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5221 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5281 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5341 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5401 tccccgaaaa gtgccacctg acgt
//
