LOCUS       sequence-176893-        6468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Dsred-tagged human Melanophilin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6468)
  AUTHORS   Westbroek W, Tuchman M, Tinloy B, De Wever O, Vilboux T, Hertz JM, 
            Hasle H, Heilmann C, Helip-Wooley A, Kleta R, Gahl WA
  TITLE     A novel missense mutation (G43S) in the switch I region of Rab27A 
            causing Griscelli syndrome.
  JOURNAL   Mol Genet Metab. 2008 Jun;94(2):248-54. doi: 
            10.1016/j.ymgme.2008.02.009. Epub 2008 Apr 7.
  PUBMED    18397837
REFERENCE   2  (bases 1 to 6468)
  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"; doi: 
            "10.1016/j.ymgme.2008.02.009"; journalName: "Mol Genet Metab"; date:
            "2008-06"; volume: "94"; issue: "2"; pages: "248-54"
COMMENT     Sequence Label: pDsred monomer C1- MLPH
FEATURES             Location/Qualifiers
     source          1..6468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        779..1082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1083..1286
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1237..1257
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1325..1334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1331..2005
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     primer_bind     1729..1748
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3974..4095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4011..4030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4065..4084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4102..4557)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4239..4260)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4451..4470
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4584..4688
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4690..5047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4690..4881
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4713..4733)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4898..5033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5082..5876
                     /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             5082..5876
                     /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(5136..5155)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5746..5765
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6064..6083)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6108..6155
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
      721 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      781 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      841 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      901 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      961 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1021 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1081 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     1141 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1201 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1261 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     1321 ggtcgccacc atggacaaca ccgaggacgt catcaaggag ttcatgcagt tcaaggtgcg
     1381 catggagggc tccgtgaacg gccactactt cgagatcgag ggcgagggcg agggcaagcc
     1441 ctacgagggc acccagaccg ccaagctgca ggtgaccaag ggcggccccc tgcccttcgc
     1501 ctgggacatc ctgtcccccc agttccagta cggctccaag gcctacgtga agcaccccgc
     1561 cgacatcccc gactacatga agctgtcctt ccccgagggc ttcacctggg agcgctccat
     1621 gaacttcgag gacggcggcg tggtggaggt gcagcaggac tcctccctgc aggacggcac
     1681 cttcatctac aaggtgaagt tcaagggcgt gaacttcccc gccgacggcc ccgtaatgca
     1741 gaagaagact gccggctggg agccctccac cgagaagctg tacccccagg acggcgtgct
     1801 gaagggcgag atctcccacg ccctgaagct gaaggacggc ggccactaca cctgcgactt
     1861 caagaccgtg tacaaggcca agaagcccgt gcagctgccc ggcaaccact acgtggactc
     1921 caagctggac atcaccaacc acaacgagga ctacaccgtg gtggagcagt acgagcacgc
     1981 cgaggcccgc cactccggct cccagtccgg actcagatct cgagctcaag cttcgaattc
     2041 aatggggaag aaactggatc tttccaagct cactgatgaa gaggcccagc atgtcttgga
     2101 agttgttcaa cgagattttg acctccgaag gaaagaagag gaacggctag aggcgttgaa
     2161 gggcaagatt aagaaggaaa gctccaagag ggagctgctt tccgacactg cccatctgaa
     2221 cgagacccac tgcgcccgct gcctgcagcc ctaccagctg cttgtgaata gcaaaaggca
     2281 gtgcctggaa tgtggcctct tcacctgcaa aagctgtggc cgcgtccacc cggaggagca
     2341 gggctggatc tgtgacccct gccatctggc cagagtcgtg aagatcggct cactggagtg
     2401 gtactatgag catgtgaaag cccgcttcaa gaggttcgga agtgccaagg tcatccggtc
     2461 cctccacggg cggctgcagg gtggagctgg gcctgaactg atatctgaag agagaagtgg
     2521 agacagcgac cagacagatg aggatggaga acctggctca gaggcccagg cccaggccca
     2581 gccctttggc agcaaaaaaa agcgcctcct ctccgtccac gacttcgact tcgagggaga
     2641 ctcagatgac tccactcagc ctcaaggtca ctccctgcac ctgtcctcag tccctgaggc
     2701 cagggacagc ccacagtccc tcacagatga gtcctgctca gagaaggcag cccctcacaa
     2761 ggctgagggc ctggaggagg ctgatactgg ggcctctggg tgccactccc atccggaaga
     2821 gcagccgacc agcatctcac cttccagaca cggcgccctg gctgagctct gcccgcctgg
     2881 aggctcccac aggatggccc tggggactgc tgctgcactc gggtcgaatg tcatcaggaa
     2941 tgagcagctg cccctgcagt acttggccga tgtggacacc tctgatgagg aaagcatccg
     3001 ggctcacgtg atggcctccc accattccaa gcggagaggc cgggcgtctt ctgagagtca
     3061 gatctttgag ctgaataagc atatttcagc tgtggaatgc ctgctgacct acctggagaa
     3121 cacagttgtg cctcccttgg ccaagggtct aggtgctgga gtgcgcacgg aggccgatgt
     3181 agaggaggag gccctgagga ggaagctgga ggagctgacc agcaacgtca gtgaccagga
     3241 gacctcgtcc gaggaggagg aagccaagga cgaaaaggca gagcccaaca gggacaaatc
     3301 agttgggcct ctcccccagg cggacccgga ggtgggcacg gctgcccatc aaaccaacag
     3361 acaggaaaaa agcccccagg accctgggga ccccgtccag tacaacagga ccacagatga
     3421 ggagctgtca gagctggagg acagagtggc agtgacggcc tcagaagtcc agcaggcaga
     3481 gagcgaggtt tcagacattg aatccaggat tgcagccctg agggccgcag ggctcacggt
     3541 gaagccctcg ggaaagcccc ggaggaagtc aaacctcccg atatttctcc ctcgagtggc
     3601 tgggaaactt ggcaagagac cagaggaccc aaatgcagac ccttcaagtg aggccaaggc
     3661 aatggctgtg ccctatcttc tgagaagaaa gttcagtaat tccctgaaaa gtcaaggtaa
     3721 agatgatgat tcttttgatc ggaaatcagt gtaccgaggc tcgctgacac agagaaaccc
     3781 caacgcgagg aaaggaatgg ccagccacac cttcgcgaaa cctgtggtgg cccaccagtc
     3841 ctaaggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3901 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3961 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4021 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4081 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4141 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4201 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4261 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4321 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4381 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4441 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4501 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4561 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4621 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4681 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4741 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4801 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4861 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4921 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4981 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5041 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5101 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5161 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5221 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5281 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5341 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5401 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5461 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5521 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5581 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5641 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5701 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5761 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5821 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5881 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5941 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6001 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6061 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6121 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6181 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6241 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6301 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6361 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6421 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaa
//
