LOCUS       sequence-394261-        3550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini circle middle entry plasmid for membrane mCherry-pA with 
            flanking BsaI cut sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3550)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3550)
  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            "Unpublished"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_memb-mCherry-pA
FEATURES             Location/Qualifiers
     source          1..3550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(16..102)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             326..1036
                     /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(471..489)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     739..758
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1074..1208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1098..1117)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1152..1171
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1645..2436
                     /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      2529..3117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3018..3037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3271..3288
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3447..3474)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
       61 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      121 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtcttaagct cgggcccggt ctctagtcgc caccatgctg tgctgcatca gaagaactaa
      241 accggttgag aagaatgaag aggccgatca ggagctgcag tcgacggtac cgcgggcccg
      301 ggacccaccg gtcgccacca agcttatggt gagcaagggc gaggaggata acatggccat
      361 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
      421 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      481 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
      541 cggctccaag gcctacgtga agcacccagc cgacatcccc gactacttga agctgtcctt
      601 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
      661 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
      721 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
      781 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
      841 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
      901 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
      961 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     1021 cgagctgtac aagtagctcg agcctctaga actatagtga gtcgtattac gtagatccag
     1081 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1141 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1201 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1261 aggtttttta attcctcaag agacctctag accagccagg acagaaatgc ctcgacttcg
     1321 ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag
     1381 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
     1441 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     1501 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
     1561 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
     1621 cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa
     1681 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
     1741 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
     1801 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
     1861 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
     1921 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
     1981 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
     2041 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
     2101 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
     2161 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     2221 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     2281 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     2341 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     2401 gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa
     2461 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2641 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3181 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3241 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3301 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3361 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     3421 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3481 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     3541 caacgttcaa
//
