LOCUS       sequence-482175-        3598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector for mini-Golden subcloning platform; mNeongreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3598)
  TITLE     Mini-Golden Subcloning Platform
REFERENCE   2  (bases 1 to 3598)
  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"
COMMENT     Sequence Label: SA-memb-mNeongreen
FEATURES             Location/Qualifiers
     source          1..3598
                     /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"
     CDS             367..1077
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1107..1241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1131..1150)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1185..1204
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1678..2469
                     /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      2562..3150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3051..3070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3304..3321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3480..3507)
                     /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 ccggtctctt cgccactaat ccggagtgtg tgtgtgtttc tgacgtgtct cttccctttt
      241 ttttttcagg ccaccatgct gtgctgcatc agaagaacta aaccggttga gaagaatgaa
      301 gaggccgatc aggagctgca gtcgacggta ccgcgggccc gggacccacc ggtcgccacc
      361 aagcttatgg tgagcaaggg cgaggaggat aacatggcct ctctcccagc gacacatgag
      421 ttacacatct ttggctccat caacggtgtg gactttgaca tggtgggtca gggcaccggc
      481 aatccaaatg atggttatga ggagttaaac ctgaagtcca ccaagggtga cctccagttc
      541 tccccctgga ttctggtccc tcatatcggg tatggcttcc atcagtacct gccctaccct
      601 gacgggatgt cgcctttcca ggccgccatg gtagatggct ccggctacca agtccatcgc
      661 acaatgcagt ttgaagatgg tgcctccctt actgttaact accgctacac ctacgaggga
      721 agccacatca aaggagaggc ccaggtgaag gggactggtt tccctgctga cggtcctgtg
      781 atgaccaact cgctgaccgc tgcggactgg tgcaggtcga agaagactta ccccaacgac
      841 aaaaccatca tcagtacctt taagtggagt tacaccactg gaaatggcaa gcgctaccgg
      901 agcactgcgc ggaccaccta cacctttgcc aagccaatgg cggctaacta tctgaagaac
      961 cagccgatgt acgtgttccg taagacggag ctcaagcact ccaagaccga gctcaacttc
     1021 aaggagtggc aaaaggcctt taccgatgtg atgggcatgg acgagctgta caagtaagct
     1081 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
     1141 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1201 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1261 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcctc aagagacctc
     1321 tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
     1381 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
     1441 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     1501 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
     1561 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     1621 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
     1681 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
     1741 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
     1801 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
     1861 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
     1921 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
     1981 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
     2041 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
     2101 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
     2161 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
     2221 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     2281 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     2341 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     2401 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     2461 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     2521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2701 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3181 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3241 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3301 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3361 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3421 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     3481 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     3541 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcgg
//
