LOCUS       sequence-394487-        3425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini circle middle entry plasmid for BFP-pA with flanking BsaI cut 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3425)
  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            "Unpublished"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-pA
FEATURES             Location/Qualifiers
     source          1..3425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..26
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(185..212)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(304..390)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     439..461
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     453..470
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     454..470
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             503..1204
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1237..1371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1261..1280)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1315..1334
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1808..2599
                     /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      2692..3280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3181..3200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
       61 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      121 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      181 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      241 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      301 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      361 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      421 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      481 ggcccggtct ctagtcgcca ccatgagcga gctgattaag gagaacatgc acatgaagct
      541 gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg gcgaaggcaa
      601 gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt
      661 cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca tcaaccacac
      721 ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat gggagagagt
      781 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
      841 ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg gccctgtgat
      901 gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg ctgacggcgg
      961 cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa
     1021 catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta
     1081 ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacgt acgtcgagca
     1141 gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc acaagcttaa
     1201 ttaataagct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
     1261 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1321 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1381 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcctc
     1441 aagagacctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc
     1501 cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc
     1561 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     1621 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     1681 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     1741 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     1801 aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     1861 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     1921 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
     1981 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     2041 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     2101 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     2161 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     2221 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     2281 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     2341 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     2401 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     2461 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     2521 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     2581 caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta
     2641 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2701 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2761 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2821 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2881 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2941 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3001 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3061 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3121 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3181 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3241 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3301 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3361 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3421 aacga
//
