LOCUS       sequence-446958-        3653 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 3653)
  AUTHORS   Bryksin AV, Matsumura I
  TITLE     Rational design of a plasmid origin that replicates efficiently in 
            both gram-positive and gram-negative bacteria.
  JOURNAL   PLoS One. 2010  . 5(10):e13244.
  PUBMED    20949038
REFERENCE   2  (bases 1 to 3653)
  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: "PLoS One. 
            2010  . 5(10):e13244."
COMMENT     Sequence Label: pBAV1K-T5-gfp
FEATURES             Location/Qualifiers
     source          1..3653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(26..40,41..46,47..63,64..70)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    46..62
                     /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)."
     protein_bind    78..94
                     /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)."
     RBS             104..115
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(152..180)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     775..795
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    842..862
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      913..999
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2330..2341
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(2543..2637)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2653..3447)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    join(3653,1..21)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aattcgcggc cgcttctaga ggaaatcata aaaaatttat ttgctttgtg agcggataac
       61 aattataata gattcaattg tgagcggata acaattacta gagaaagagg agaaatacta
      121 gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
      181 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
      241 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
      301 tgtcactact ctgacgtatg gtgttcaatg cttttcccgt tatccggatc atatgaaacg
      361 gtatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
      421 caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
      481 taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa
      541 actcgagtac aactataact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
      601 aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggatccgttc aactagcaga
      661 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
      721 cctgtcgaca caatctgccc ttttgaaaga tcccaacgaa aagcgtgacc acatggtcct
      781 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgaactat acaaataata
      841 atactagtag cggccgctgc aggcctcagg gcccgatcga tgccgccgct taattaatta
      901 atccagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      961 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagtgt
     1021 cattttattt cccccgtttc agcatcaaga acctttgcat aacttgctct atatccacac
     1081 tgataattgc cctcaaacca taatctaaag gcgctagagt ttgttgaaac aatatctttt
     1141 acatcattcg tatttaaaat tccaaactcc gctcccctaa ggcgaataaa agccattaaa
     1201 tcttttgtat ttaccaaatt atagtcatcc actatatcta agagtaaatt cttcaattct
     1261 cttttttggc tttcatcaag tgttatatag cggtcaatat caaaatcatt aatgttcaaa
     1321 atatcttttt tgtcgtatat atgtttattc ttagcaatag cgtcctttga ttcatgagtc
     1381 aaatattcat atgaaccttt gatataatca agtatctcaa catgagcaac tgaactattc
     1441 cccaattttc gcttaatctt gttcctaacg ctttctattg ttacaggatt tcgtgcaata
     1501 tatataacgt gatagtgtgg ttttttatag tgctttccat ttcgtataac atcactacta
     1561 ttccatgtat ctttatcttt tttttcgtcc atatcgtgta aaggactgac agccatagat
     1621 acgcccaaac tctctaattt ttccttccaa tcattaggaa ttgagtcagg atataataaa
     1681 aatccaaaat ttctagcttt agtattttta atagccatga tataattacc ttatcaaaaa
     1741 caagtagcga aaactcgtat ccttctaaaa acgcgagctt tcgcttattt tttttgttct
     1801 gattcctttc ttgcatattc ttctatagct aacgccgcaa ccgcagattt tgaaaaacct
     1861 ttttgtttcg ccatatctgt taatttttta tcttgctctt ttgtcagaga aatcataact
     1921 ctttttttcg attctgaaat caccatttaa aaaactccaa tcaaataatt ttataaagtt
     1981 agtgtatcac tttgtaatca taaaaacaac aataaagcta cttaaatata gatttataaa
     2041 aaacgttggc gaaaacgttg gcgattcgtt ggcgattgaa aaacccctta aacccttgag
     2101 ccagttggga tagagcgttt ttggcacaaa aattggcact cggcacttaa tggggggtcg
     2161 tagtacggaa gcaaaattcg cttcctttcc ccccattttt ttccaaattc cagatttttt
     2221 tcaaaaattt tccagcgcta ccgctcggca aaattgcaag caatttttaa aatcaaaccc
     2281 atgagggaat ttcattccct catactccct tgagcctcct ccaaccgaaa tagaagggcg
     2341 ctgcgcttat tatttcattc agtcatcggc tttcataatc taacagacaa catcttcgct
     2401 gcaaagccac gctacgctca agggctttta cgctacgata acgcctgttt taacgattat
     2461 gccgataact aaacgaaata aacgctaaaa cgtctcagaa acgattttga gacgttttaa
     2521 taaaaaatcg cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     2581 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac gggagtccaa
     2641 gcgagctcgg tactaaaaca attcatccag taaaatataa tattttattt tctcccaatc
     2701 aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc
     2761 cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc
     2821 aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc
     2881 gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc
     2941 gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc
     3001 gttattcagt aagtaatcca attcggctaa gcggccgtct aagctattcg tatagggaca
     3061 atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct cgatagtctt
     3121 ttcagggctt tgttcatctt catacccttc cgagcaaagg acgccatcgg cctcactcat
     3181 gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt
     3241 tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata
     3301 ccggctgtcc gtcattttta aatataggat ttcattttct cccaccagct tatatacctt
     3361 agcaggagac attccttccg tatcttttac gcagcggtat tcttcgatca gttttttcaa
     3421 ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct acagtattta
     3481 aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc
     3541 taaaacctta aatacagaaa acagcctttt caaagttgtt ttcaaagttg gcgtataaca
     3601 tagtatcgac ggagccgatt ttgaaaccac aattatgata gaatttgacg tcg
//
