LOCUS       sequence-8298-ex        7916 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 7916)
  AUTHORS   Luong TT, Lee CY
  TITLE     Improved single-copy integration vectors for Staphylococcus aureus.
  JOURNAL   J Microbiol Methods. 2007 Jul . 70(1):186-90.
  PUBMED    17512993
REFERENCE   2  (bases 1 to 7916)
  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: "J Microbiol
            Methods. 2007 Jul . 70(1):186-90."
COMMENT     Sequence Label: pLL2787
FEATURES             Location/Qualifiers
     source          1..7916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1375..2457
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1394..1413)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        2712..2816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2817..2885,2886..3677)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3035..3054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(3856..3878)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3964..4104
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4019..4038
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4370..4389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6208..7212)
                     /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"
     protein_bind    7898..7914
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 aagcttgcat gcctgcagcc aattttgagg agggagaagc aaaatgccag tatataagga
       61 tgataataca ggtaaatggt atttttccat tagatataaa gatgtatacg gtaataacaa
      121 acgaaaaatg aagcgtgggt ttgaacgtaa gaaagatgcc aaactagctg aaagcgaatt
      181 tatacaaaat gttaaatatg gatactcgga caatcaaccc tttgaatata tattttttga
      241 tcgtttaaaa aatgaaaatc tttctgcacg ctcaatagaa aagcgaacta cagaatataa
      301 tactcacata aaagaaaggt tcggaaatat ccctattggc aaaatcacta ctacgcaatg
      361 tactgctttc aggaattatt tgttaaacga tgcaggtctt tctgttgact atgcacgatc
      421 tgtgtgggca ggttttaaag cagttatcaa ttacgccaaa aagcattaca agctcttata
      481 cgaccccaca ttatcggtaa ctcctattcc cagaacaaaa ccacaagcta aatttatcac
      541 tcgtgaagaa tttgatgaaa aagtagaaca aatcacaaat gatacttctc gtcagctaac
      601 tagactgtta ttttattctg gtcttagaat aggagaagct ttagctttgc agtggaaaga
      661 ttacgataaa ataaaaggcg aaattgacgt aaataagaaa atcaatttaa gtaatagaaa
      721 aattgaatat aatctaaaaa aagaaagctc taaagggata atacctgtac caaatttaat
      781 tagagagatg cttaaaaaca tgtataatga atcttctaaa agatataaat attttgacga
      841 aaactatttt atattcgggg gtttagaacc tattagatac gttacttatt cgtatcattt
      901 taaatctgta ttcccgaatc taaaaataca ccatttaaga cactcgtacg ctagctattt
      961 aattaataat ggtgtagata tgtatttatt aatggaatta atgaggcatt ctaacattac
     1021 agaaacaatt caaacgtact ctcatttata tactgataaa aaacatcaag ctatgagcat
     1081 atttgattaa acggtatcaa attggatccc cgggtaccga gctcgaattc ccaagctaat
     1141 tcggtggaaa cgaggtcatc atttccttcc gaaaaaacgg ttgcatttaa atcttacata
     1201 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgagtcggct gaaagatcgt
     1261 acgtaccaat tattgtttcg tgattgttca agccataaca ctgtagggat agtggaaaga
     1321 gtgcttcatc tggttacgat caatcaaata ttcaaacgga gggagacgat tttgatgaaa
     1381 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     1441 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     1501 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     1561 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     1621 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     1681 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     1741 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     1801 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     1861 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     1921 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     1981 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     2041 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     2101 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     2161 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     2221 atcccgccgt caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     2281 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     2341 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2401 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2461 caacgcaatt aatgtgagtt aggatctcac atgttctttc ctgcgttatc ccctgattct
     2521 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2581 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     2641 cccgcgcgtt ggccgattca ttaatgcaga ttaacgctta caatttaggt ggcacttttc
     2701 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2761 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3001 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3361 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3781 aaatccctta acgtgagttt tcgttccact gagcgtcaga tcctgcctcg cgcgtttcgg
     3841 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3901 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3961 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     4021 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4081 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatct
     4801 ggagctgtaa tataaaaacc ttcttcaact aacggggcag gttagtgaca ttagaaaacc
     4861 gactgtaaaa agtacagtcg gcattatctc atattataaa agccagtcat taggcctatc
     4921 tgacaattcc tgaatagagt tcataaacaa tcctgcatga taaccatcac aaacagaatg
     4981 atgtacctgt aaagatagcg gtaaatatat tgaattacct ttattaatga attttcctgc
     5041 tgtaataatg ggtagaaggt aattactatt attattgata tttaagttaa acccagtaaa
     5101 tgaagtccat ggaataatag aaagagaaaa agcattttca ggtataggtg ttttgggaaa
     5161 caatttcccc gaaccattat atttctctac atcagaaagg tataaatcat aaaactcttt
     5221 gaagtcattc tttacaggag tccaaatacc agagaatgtt ttagatacac catcaaaaat
     5281 tgtataaagt ggctctaact tatcccaata acctaactct ccgtcgctat tgtaaccagt
     5341 tctaaaagct gtatttgagt ttatcaccct tgtcactaag aaaataaatg cagggtaaaa
     5401 tttatatcct tcttgtttta tgtttcggta taaaacacta atatcaattt ctgtggttat
     5461 actaaaagtc gtttgttggt tcaaataatg attaaatatc tcttttctct tccaattgtc
     5521 taaatcaatt ttattaaagt tcatttgata tgcctcctaa atttttatct aaagtgaatt
     5581 taggaggctt acttgtctgc tttcttcatt agaatcaatc cttttttaaa agtcaatatt
     5641 actgtaacat aaatatatat tttaaaaata tcccacttta tccaattttc gtttgttgaa
     5701 ctaatgggtg ctttagttga agaataaaga ccacattaaa aaatgtggtc ttttgtgttt
     5761 ttttaaagga tttgagcgta gcgaaaaatc cttttctttc ttatcttgat aataagggta
     5821 actattgccg gttgtccatt catggctgaa ctctgcttcc tctgttgaca tgacacacat
     5881 catctcaata tccgaatagg gcccatcagt ctgacgacca agagagccat aaacaccaat
     5941 agccttaaca tcatccccat atttatccaa tattcgttcc ttaatttcat gaacaatctt
     6001 cattctttct tctctagtca ttattattgg tccattcact attctcattc ccttttcaga
     6061 taattttaga tttgcttttc taaataagaa tatttggaga gcaccgttct tattcagcta
     6121 ttaataactc gtcttcctaa gcatccttca atccttttaa taacaattat agcatctaat
     6181 cttcaacaaa ctggcccgtt tgttgaacta ctctttaata aaataatttt tccgttccca
     6241 attccacatt gcaataatag aaaatccatc ttcatcggct ttttcgtcat catctgtatg
     6301 aatcaaatcg ccttcttctg tgtcatcaag gtttaatttt ttatgtattt cttttaacaa
     6361 accaccatag gagattaacc ttttacggtg taaaccttcc tccaaatcag acaaacgttt
     6421 caaattcttt tcttcatcat cggtcataaa atccgtatcc tttacaggat attttgcagt
     6481 ttcgtcaatt gccgattgta tatccgattt atatttattt ttcggtcgaa tcatttgaac
     6541 ttttacattt ggatcatagt ctaatttcat tgcctttttc caaaattgaa tccattgttt
     6601 ttgattcacg tagttttctg tattcttaaa ataagttggt tccacacata ccaatacatg
     6661 catgtgctga ttataagaat tatctttatt atttattgtc acttccgttg cacgcataaa
     6721 accaacaaga tttttattaa tttttttata ttgcatcatt cggcgaaatc cttgagccat
     6781 atctgacaaa ctcttattta attcttcgcc atcataaaca tttttaactg ttaatgtgag
     6841 aaacaaccaa cgaactgttg gcttttgttt aataacttca gcaacaacct tttgtgactg
     6901 aatgccatgt ttcattgctc tcctccagtt gcacattgga caaagcctgg atttacaaaa
     6961 ccacactcga tacaactttc tttcgcctgt ttcacgattt tgtttatact ctaatatttc
     7021 agcacaatct tttactcttt cagccttttt aaattcaaga atatgcagaa gttcaaagta
     7081 atcaacatta gcgattttct tttctctcca tggtctcact tttccacttt ttgtcttgtc
     7141 cactaaaacc cttgattttt catctgaata aatgctacta ttaggacaca taatattaaa
     7201 agaaaccccc atctatttag ttatttgttt agtcacttat aactttaaca gatggggttt
     7261 ttctgtgcaa ccaattttaa gggttttcaa tactttaaaa cacatacata ccaacacttc
     7321 aacgcacctt tcagcaacta aaataaaaat gacgttattt ctatatgtat caagataaga
     7381 aagaacaagt tcaaaaccat caaaaaaaga caccttttca ggtgcttttt ttattttata
     7441 aactcattcc ctgatctcga cttcgttctt tttttacctc tcggttatga gttagttcaa
     7501 attcgttctt tttaggttct aaatcgtgtt tttcttggaa ttgtgctgtt ttatccttta
     7561 ccttgtctac aaacccctta aaaacgtttt taaaggcttt taagccgtct gtacgttcct
     7621 taaggaatta attctacaca gcccagtcca gactattcgg cactgaaatt atgggtgaag
     7681 tggtcaagac ctcactaggc accttaaaaa tagcgcaccc tgaagaagat ttatttgagg
     7741 tagcccttgc ctacctagct tccaagaaag atatcctaac agcacaagag cggaaagatg
     7801 ttttgttcta catccagaac aacctctgct aaaattcctg aaaaattttg caaaaagttg
     7861 ttgactttat ctacaaggtg tggcataatg tgtggaattg tgagcggata acaatt
//
