LOCUS       sequence-425590-        6439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for transient transfection of RDX-mCherry fusion protein in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6439)
  AUTHORS   Miller SG, Hoh M, Ebmeier CC, Tay JW, Ahn NG
  TITLE     Cooperative polarization of MCAM/CD146 and ERM family proteins in 
            melanoma.
  JOURNAL   Mol Biol Cell. 2023 Dec 20:mbcE23060255. doi: 
            10.1091/mbc.E23-06-0255.
  PUBMED    38117590
REFERENCE   2  (bases 1 to 6439)
  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: "Mol Biol 
            Cell. 2023 Dec 20:mbcE23060255. doi: 10.1091/mbc.E23-06-0255."
COMMENT     Sequence Label: RDX-mCherry
FEATURES             Location/Qualifiers
     source          1..6439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2480..2489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2486..3196
                     /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(2631..2649)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2899..2918
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3319..3440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3356..3375)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3410..3429
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3447..3902)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3584..3605)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3796..3815
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3929..4033
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4035..4392
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4035..4226
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4058..4078)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4243..4378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4427..5221
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4427..5221
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4481..4500)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5091..5110
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5409..5428)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5453..5500
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5829..6417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6318..6337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 ggtttagtga accgtcagat ccgctagcgc caccatgccg aaaccaatca acgtaagagt
      721 aactacaatg gatgctgagc tggaatttgc cattcagccc aatacaactg gcaaacaact
      781 ttttgaccag gtggtgaaaa cagttggttt gcgtgaggtc tggttttttg ggctgcagta
      841 tgtagacagc aaaggttatt ctacatggct taaactaaat aaaaaggtaa cacagcagga
      901 tgttaaaaaa gagaatcctt tacagttcaa gtttagagct aaattctttc ctgaagatgt
      961 ttctgaggaa ttaattcaag aaataaccca gagactcttc ttcttgcaag ttaaagaagc
     1021 catcttaaat gatgagatat attgcccgcc agaaactgca gttcttttgg cttcctatgc
     1081 tgtccaagcc aagtatggag attacaataa agagattcat aagccaggct acctggctaa
     1141 tgatagactc ctaccccagc gtgtattgga acaacacaaa ctaacaaaag aacagtggga
     1201 agaaagaata cagaactggc atgaagaaca tagaggaatg ttaagggagg attctatgat
     1261 ggaatacctg aagattgcac aagatctaga aatgtatgga gtcaactatt ttgaaataaa
     1321 aaataaaaaa ggaactgaat tgtggctagg tgttgatgct ttgggtctga atatttatga
     1381 gcatgacgac aagttaacac ctaaaattgg ttttccctgg agtgaaatca gaaatatttc
     1441 atttaatgac aaaaaatttg ttataaagcc aatcgacaaa aaggcacctg attttgtgtt
     1501 ttatgcacct cgtctgagaa tcaataagag gattttggcc ttatgtatgg gaaaccatga
     1561 actatacatg cgaagaagga agcctgatac tattgaagta caacagatga aggctcaggc
     1621 tagggaggag aaacatcaga agcagttgga aagggcacaa ttagagaatg aaaaggagaa
     1681 aagagaaata gcagaaaagg aaaaggaaag aatagaacgt gaaaaggaag agctaatgga
     1741 acgtctaaaa caaattgaag agcagacaat taaagctcag aaagaactag aagaacagac
     1801 tcgaaaagct ctagaactgg atcaagaacg aaaacgagca aaagaagaag cagaacgact
     1861 tgaaaaggag cgtcgagctg ctgaagaggc aaagtctgcc atagcaaaac aagctgccga
     1921 ccagatgaag aatcaggagc agctagcagc agaacttgct gaattcactg ccaagattgc
     1981 acttctagag gaagccaaga agaaaaagga agaggaagct actgagtggc aacacaaagc
     2041 ttttgcagcc caggaagact tggaaaagac caaagaagag ttaaaaactg tgatgtctgc
     2101 cccccctcca cctccaccac caccagtcat tcctccaaca gaaaacgaac atgatgaaca
     2161 cgatgagaat aatgctgaag ctagtgctga attatcaaat gaaggggtaa tgaaccatag
     2221 aagcgaggaa gaacgtgtaa ccgaaacaca gaaaaatgag cgtgttaaga agcaacttca
     2281 ggcattaagt tcagaattag cccaagccag agatgaaacc aagaaaacac aaaatgatgt
     2341 tcttcatgct gagaatgtta aagcaggccg tgataagtac aagactctgc gacagattcg
     2401 acaaggcaat acaaagcagc gtatcgatga gtttgaagca atggcatccg gcggtagcgg
     2461 cggtggcagc ggaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat
     2521 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     2581 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     2641 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     2701 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     2761 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     2821 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     2881 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     2941 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     3001 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     3061 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     3121 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     3181 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3241 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3301 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3361 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3421 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3481 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3541 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3601 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3661 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3721 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3781 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3841 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3901 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3961 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4021 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4081 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4141 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4201 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4261 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4321 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4381 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4441 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4501 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4561 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4621 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4681 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4741 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4801 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4861 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4921 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4981 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5041 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5101 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5161 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5221 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5281 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5341 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5401 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5461 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5521 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5581 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5641 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5941 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6421 gccagcaacg cggcctttt
//
