LOCUS       sequence-394644-        4145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini circle middle entry plasmid for BFP-P2A-NTR version 2-pA with 
            flanking BsaI cut sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4145)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4145)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMC-ME_BFP-P2A-NTRv2-pA
FEATURES             Location/Qualifiers
     source          1..4145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(12..98)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     147..169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     161..178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     162..178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             211..909
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             919..975
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    1665..1799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1689..1708)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1743..1762
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2236..3027
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3120..3708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3609..3628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3862..3879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4038..4065)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
       61 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      121 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct
      181 taagctcggg cccggtctct agtcgccacc atgagcgagc tgattaagga gaacatgcac
      241 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
      301 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
      361 ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc
      421 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
      481 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
      541 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
      601 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
      661 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
      721 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
      781 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacgtac
      841 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
      901 aagcttaatg gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag
      961 gagaaccccg ggcctatgac tattgttcaa gctgcccaat cccgctactc caccaaagcc
     1021 tttgatgctt cgcgcaaatt gcctgaagag aaagtcgcgg cagtgaaaga gttaatccgc
     1081 atgagtgcgt ccagtgtcaa ctcgcaacct tggcatttta ttgtcgcgag cagtgaagag
     1141 ggaaaagcgc gcatcgccaa agcaacacaa ggtggttttg ctgctaatga gcgcaagatt
     1201 ttggatgctt ctcacgttgt ggtgttctgc gccaaaacgg cgattgatga agcgtaccta
     1261 ctcgaccttt tggaaagcga agacaaagat ggccgctacg ccgatgtcga agcaaaaaat
     1321 ggcatgcacg ctggtcgttc atttttcgtc aacatgcacc gctttgactt gaaagacgcg
     1381 catcactgga tggaaaagca agtttacctc aatgtgggga cgctgctatt gggtgcttct
     1441 gcgatggaga tcgacgcggt gccaattgaa ggcttcgatg ccaaggtgct tgatgaggag
     1501 tttggtctgc gtgagaaggg ctttaccagc gtggtgattg tgccgctggg ttaccatagc
     1561 gaagacgatt ttaatgctaa gctgccaaaa tcacgttggt cagcagagac tgtttttacc
     1621 gaaatctaac tcgagtctag aactatagtg agtcgtatta cgtagatcca gacatgataa
     1681 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1741 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1801 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1861 aattcctcaa gagacctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc
     1921 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa
     1981 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     2041 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     2101 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     2161 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     2221 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     2281 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     2341 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     2401 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     2461 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     2521 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     2581 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     2641 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     2701 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     2761 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     2821 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     2881 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     2941 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     3001 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac
     3061 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3121 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3181 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3241 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3301 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3361 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3421 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3481 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3541 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3601 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3661 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3721 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3781 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3841 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3901 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3961 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     4021 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     4081 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     4141 aatcc
//
