LOCUS       sequence-394482-        4154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini circle middle entry plasmid for mCherry- 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 4154)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4154)
  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            "Unpublished"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 mCherry-P2A-NTRv2-pA
FEATURES             Location/Qualifiers
     source          1..4154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..87)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      194..203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             200..907
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(345..363)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     613..632
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     807..830
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             917..973
                     /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    1663..1797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1687..1706)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1741..1760
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2234..3025
                     /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      3118..3706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3607..3626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3860..3877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4036..4063)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      121 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      181 ccggtctcta gtcgccacca tggtgagcaa gggcgaggag gacaacatgg ccatcatcaa
      241 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
      301 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      361 caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
      421 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
      481 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      541 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
      601 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
      661 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
      721 cggcggccac tacgacgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
      781 gcccggcgcc tacaacgtca acatcaagct ggacatcacc tcccacaacg aggactacac
      841 catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcggca tggacgagct
      901 gtacaaggga agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga
      961 gaaccccggg cctatgacta ttgttcaagc tgcccaatcc cgctactcca ccaaagcctt
     1021 tgatgcttcg cgcaaattgc ctgaagagaa agtcgcggca gtgaaagagt taatccgcat
     1081 gagtgcgtcc agtgtcaact cgcaaccttg gcattttatt gtcgcgagca gtgaagaggg
     1141 aaaagcgcgc atcgccaaag caacacaagg tggttttgct gctaatgagc gcaagatttt
     1201 ggatgcttct cacgttgtgg tgttctgcgc caaaacggcg attgatgaag cgtacctact
     1261 cgaccttttg gaaagcgaag acaaagatgg ccgctacgcc gatgtcgaag caaaaaatgg
     1321 catgcacgct ggtcgttcat ttttcgtcaa catgcaccgc tttgacttga aagacgcgca
     1381 tcactggatg gaaaagcaag tttacctcaa tgtggggacg ctgctattgg gtgcttctgc
     1441 gatggagatc gacgcggtgc caattgaagg cttcgatgcc aaggtgcttg atgaggagtt
     1501 tggtctgcgt gagaagggct ttaccagcgt ggtgattgtg ccgctgggtt accatagcga
     1561 agacgatttt aatgctaagc tgccaaaatc acgttggtca gcagagactg tttttaccga
     1621 aatctaactc gagtctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
     1681 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1741 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1801 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1861 ttcctcaaga gacctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     1921 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     1981 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     2041 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     2101 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     2161 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     2221 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     2281 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     2341 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     2401 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     2461 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     2521 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     2581 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     2641 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     2701 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     2761 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     2821 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     2881 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     2941 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     3001 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt
     3061 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3481 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3541 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3601 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3661 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3721 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3781 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3841 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3901 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3961 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     4021 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     4081 gcagtttatg gcgggcgtcc tgcccgccmc cctccgggcc gttgcttcac aacgttcaaa
     4141 tccgctcccg gcgg
//
