LOCUS       sequence-394265-        3479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini circle middle entry plasmid for 2a-mCherry-pA ORF 1 (without 
            ATG) with flanking BsaI cut sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3479)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3479)
  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            "Unpublished"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_2a-mCherry-pA ORF-1 without ATG
FEATURES             Location/Qualifiers
     source          1..3479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..101)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     150..172
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     164..181
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     165..181
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             208..264
                     /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"
     CDS             265..972
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(407..425)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     675..694
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1002..1136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1026..1045)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1080..1099
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1573..2364
                     /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      2457..3045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2946..2965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3199..3216
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3375..3402)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
       61 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
      121 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
      181 tcttaagctc gggcccggtc tctagtcgct acaaacttct ctctgctgaa gcaggctgga
      241 gacgtggagg agaaccctgg acctgtgagc aagggcgagg aggataacat ggccatcatc
      301 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      361 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      421 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
      481 tccaaggcct acgtgaagca cccagccgac atccccgact acttgaagct gtccttcccc
      541 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      601 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
      661 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
      721 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
      781 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
      841 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
      901 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
      961 ctgtacaagt aagctagaac tatagtgagt cgtattacgt agatccagac atgataagat
     1021 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1081 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1141 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     1201 tcctcaagag acctctagac cagccaggac agaaatgcct cgacttcgct gmtacccaag
     1261 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc
     1321 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
     1381 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
     1441 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1501 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     1561 aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     1621 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     1681 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
     1741 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     1801 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     1861 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     1921 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     1981 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     2041 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     2101 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     2161 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
     2221 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     2281 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
     2341 atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg
     2401 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2581 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3121 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3181 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3241 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3301 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3361 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3421 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaa
//
