LOCUS       sequence-176891-        7527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of tagged wild type human Melanophilin with Dsred monomer
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7527)
  TITLE     Dsred - monomer C1 hygro -- Human Melanophilin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7527)
  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            "Unpublished"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: pDsred monomer hygro C1- MLPH
FEATURES             Location/Qualifiers
     source          1..7527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        786..1089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1090..1293
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1244..1264
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1332..1341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1338..2012
                     /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     1736..1755
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3981..4102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4072..4091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4109..4564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4246..4267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4458..4477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4591..4695
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4697..5054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4697..4888
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4720..4740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4905..5040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5062..6087
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5062..6087
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    6204..6285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6313..6331)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6399..6503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6504..6572,6573..7364)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6722..6741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
      721 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      781 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      841 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      901 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      961 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     1021 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     1081 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     1141 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     1201 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     1261 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
     1321 cgctaccggt cgccaccatg gacaacaccg aggacgtcat caaggagttc atgcagttca
     1381 aggtgcgcat ggagggctcc gtgaacggcc actacttcga gatcgagggc gagggcgagg
     1441 gcaagcccta cgagggcacc cagaccgcca agctgcaggt gaccaagggc ggccccctgc
     1501 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggcc tacgtgaagc
     1561 accccgccga catccccgac tacatgaagc tgtccttccc cgagggcttc acctgggagc
     1621 gctccatgaa cttcgaggac ggcggcgtgg tggaggtgca gcaggactcc tccctgcagg
     1681 acggcacctt catctacaag gtgaagttca agggcgtgaa cttccccgcc gacggccccg
     1741 taatgcagaa gaagactgcc ggctgggagc cctccaccga gaagctgtac ccccaggacg
     1801 gcgtgctgaa gggcgagatc tcccacgccc tgaagctgaa ggacggcggc cactacacct
     1861 gcgacttcaa gaccgtgtac aaggccaaga agcccgtgca gctgcccggc aaccactacg
     1921 tggactccaa gctggacatc accaaccaca acgaggacta caccgtggtg gagcagtacg
     1981 agcacgccga ggcccgccac tccggctccc agtccggact cagatctcga gctcaagctt
     2041 cgaattcaat ggggaagaaa ctggatcttt ccaagctcac tgatgaagag gcccagcatg
     2101 tcttggaagt tgttcaacga gattttgacc tccgaaggaa agaagaggaa cggctagagg
     2161 cgttgaaggg caagattaag aaggaaagct ccaagaggga gctgctttcc gacactgccc
     2221 atctgaacga gacccactgc gcccgctgcc tgcagcccta ccagctgctt gtgaatagca
     2281 aaaggcagtg cctggaatgt ggcctcttca cctgcaaaag ctgtggccgc gtccacccgg
     2341 aggagcaggg ctggatctgt gacccctgcc atctggccag agtcgtgaag atcggctcac
     2401 tggagtggta ctatgagcat gtgaaagccc gcttcaagag gttcggaagt gccaaggtca
     2461 tccggtccct ccacgggcgg ctgcagggtg gagctgggcc tgaactgata tctgaagaga
     2521 gaagtggaga cagcgaccag acagatgagg atggagaacc tggctcagag gcccaggccc
     2581 aggcccagcc ctttggcagc aaaaaaaagc gcctcctctc cgtccacgac ttcgacttcg
     2641 agggagactc agatgactcc actcagcctc aaggtcactc cctgcacctg tcctcagtcc
     2701 ctgaggccag ggacagccca cagtccctca cagatgagtc ctgctcagag aaggcagccc
     2761 ctcacaaggc tgagggcctg gaggaggctg atactggggc ctctgggtgc cactcccatc
     2821 cggaagagca gccgaccagc atctcacctt ccagacacgg cgccctggct gagctctgcc
     2881 cgcctggagg ctcccacagg atggccctgg ggactgctgc tgcactcggg tcgaatgtca
     2941 tcaggaatga gcagctgccc ctgcagtact tggccgatgt ggacacctct gatgaggaaa
     3001 gcatccgggc tcacgtgatg gcctcccacc attccaagcg gagaggccgg gcgtcttctg
     3061 agagtcagat ctttgagctg aataagcata tttcagctgt ggaatgcctg ctgacctacc
     3121 tggagaacac agttgtgcct cccttggcca agggtctagg tgctggagtg cgcacggagg
     3181 ccgatgtaga ggaggaggcc ctgaggagga agctggagga gctgaccagc aacgtcagtg
     3241 accaggagac ctcgtccgag gaggaggaag ccaaggacga aaaggcagag cccaacaggg
     3301 acaaatcagt tgggcctctc ccccaggcgg acccggaggt gggcacggct gcccatcaaa
     3361 ccaacagaca ggaaaaaagc ccccaggacc ctggggaccc cgtccagtac aacaggacca
     3421 cagatgagga gctgtcagag ctggaggaca gagtggcagt gacggcctca gaagtccagc
     3481 aggcagagag cgaggtttca gacattgaat ccaggattgc agccctgagg gccgcagggc
     3541 tcacggtgaa gccctcggga aagccccgga ggaagtcaaa cctcccgata tttctccctc
     3601 gagtggctgg gaaacttggc aagagaccag aggacccaaa tgcagaccct tcaagtgagg
     3661 ccaaggcaat ggctgtgccc tatcttctga gaagaaagtt cagtaattcc ctgaaaagtc
     3721 aaggtaaaga tgatgattct tttgatcgga aatcagtgta ccgaggctcg ctgacacaga
     3781 gaaaccccaa cgcgaggaaa ggaatggcca gccacacctt cgcgaaacct gtggtggccc
     3841 accagtccta aggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     3901 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3961 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4021 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4081 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     4141 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4201 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4261 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4321 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4381 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4441 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4501 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4561 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4621 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4681 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4741 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4801 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4861 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4921 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4981 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5041 taggcttttg caaagatcga tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     5101 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     5161 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     5221 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     5281 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     5341 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     5401 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     5461 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     5521 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     5581 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     5641 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     5701 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     5761 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     5821 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     5881 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     5941 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     6001 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     6061 agcactcgtc cgagggcaaa ggaatagagt tctagaggat cataatcagc cataccacat
     6121 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6181 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcctcgc gatccacaac
     6301 catggcgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     6361 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6481 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6541 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6601 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6661 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6721 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     6781 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6841 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6901 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6961 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7021 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7081 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7141 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7201 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7261 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7321 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaggcgcg ccaggccctg
     7381 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7441 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7501 tttcgttcca ctgagcgtca gaccccg
//
