LOCUS       sequence-484620-        3509 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  Middle entry vector for mini-Golden subcloning platform; mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3509)
  TITLE     Mini-Golden Subcloning Platform
REFERENCE   2  (bases 1 to 3509)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SA-mCherry-pA
FEATURES             Location/Qualifiers
     source          1..3509
                     /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      272..281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             278..988
                     /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(423..441)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     691..710
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1018..1152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1042..1061)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1096..1115
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1589..2380
                     /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      2473..3061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2962..2981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3215..3232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3391..3418)
                     /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 ctctcttccc
      241 tttttttttt cagacggctg atttttctaa cgccaccatg gtgagcaagg gcgaggagga
      301 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      361 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      421 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      481 tcagttcatg tacggctcca aggcctacgt gaagcaccca gccgacatcc ccgactactt
      541 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      601 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
      661 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
      721 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
      781 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
      841 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
      901 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
      961 cggcggcatg gacgagctgt acaagtaagc tagaactata gtgagtcgta ttacgtagat
     1021 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1081 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1141 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1201 tgggaggttt tttaattcct caagagacct ctagaccagc caggacagaa atgcctcgac
     1261 ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     1321 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     1381 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     1441 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     1501 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     1561 cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac
     1621 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     1681 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     1741 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     1801 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     1861 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     1921 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     1981 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     2041 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     2101 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     2161 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     2221 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     2281 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     2341 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     2401 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     2461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2581 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2641 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2701 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2761 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2821 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2881 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2941 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3001 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3061 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3181 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3241 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3301 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     3361 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     3421 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     3481 ttcacaacgt tcaaatccgc tcccggcgg
//
