LOCUS       sequence-394262-        3431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mini circle middle entry plasmid for mCherry- pA with flanking BsaI 
            cut sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3431)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3431)
  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            "Unpublished"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-pA
FEATURES             Location/Qualifiers
     source          1..3431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(4..90)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     139..161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     153..170
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     154..170
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      197..206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             203..913
                     /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(348..366)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     616..635
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    943..1077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(967..986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1021..1040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1514..2305
                     /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      2398..2986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2887..2906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3140..3157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3316..3343)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
       61 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      121 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      181 ggcccggtct ctagtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat
      241 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      301 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      361 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
      421 ctccaaggcc tacgtgaagc acccagccga catccccgac tacttgaagc tgtccttccc
      481 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      541 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
      601 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
      661 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
      721 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
      781 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
      841 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
      901 gctgtacaag taagctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
      961 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1021 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1081 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1141 ttcctcaaga gacctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     1201 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     1261 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     1321 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     1381 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     1441 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     1501 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     1561 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     1621 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     1681 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     1741 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     1801 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     1861 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     1921 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     1981 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     2041 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     2101 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     2161 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     2221 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     2281 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt
     2341 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2401 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2461 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2521 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2581 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2641 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2701 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2761 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2821 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2881 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2941 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3001 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3061 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3121 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3181 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3241 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     3301 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     3361 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     3421 tccgctcccg g
//
