LOCUS       sequence-157917-        6546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Melanosome marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6546)
  AUTHORS   Ambrosio AL, Boyle JA, Aradi AE, Christian KA, Di Pietro SM
  TITLE     TPC2 controls pigmentation by regulating melanosome pH and size.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 17;113(20):5622-7. doi: 
            10.1073/pnas.1600108113. Epub 2016 May 2.
  PUBMED    27140606
REFERENCE   2  (bases 1 to 6546)
  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: 
            "10.1073/pnas.1600108113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-17- 17"; volume: "113"; issue: "20"; pages: "5622-7"
COMMENT     Sequence Label: piRFP-N3-Tyrosinase
FEATURES             Location/Qualifiers
     source          1..6546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2262..3212
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 682 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP682
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGVVIAEDRCAEVESKLGLHYPASAVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPCHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    3334..3455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3425..3444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3462..3917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3599..3620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3811..3830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3944..4048
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4050..4407
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4050..4241
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4073..4093)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4258..4393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4442..5236
                     /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             4442..5236
                     /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(4496..4515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5106..5125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5424..5443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5468..5515
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5844..6432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6333..6352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcaatgct cctggctgtt ttgtactgcc
      661 tgctgtggag tttccagacc tccgctggcc atttccctag agcctgtgtc tcctctaaga
      721 acctgatgga gaaggaatgc tgtccaccgt ggagcgggga caggagtccc tgtggccagc
      781 tttcaggcag aggttcctgt cagaatatcc ttctgtccaa tgcaccactt gggcctcaat
      841 ttcccttcac aggggtggat gaccgggagt cgtggccttc cgtcttttat aataggacct
      901 gccagtgctc tggcaacttc atgggattca actgtggaaa ctgcaagttt ggcttttggg
      961 gaccaaactg cacagagaga cgactcttgg tgagaagaaa catcttcgat ttgagtgccc
     1021 cagagaagga caaatttttt gcctacctca ctttagcaaa gcataccatc agctcagact
     1081 atgtcatccc catagggacc tatggccaaa tgaaaaatgg atcaacaccc atgtttaacg
     1141 acatcaatat ttatgacctc tttgtctgga tgcattatta tgtgtcaatg gatgcactgc
     1201 ttgggggatc tgaaatctgg agagacattg attttgccca tgaagcacca gcttttctgc
     1261 cttggcatag actcttcttg ttgcggtggg aacaagaaat ccagaagctg acaggagatg
     1321 aaaacttcac tattccatat tgggactggc gggatgcaga aaagtgtgac atttgcacag
     1381 atgagtacat gggaggtcag caccccacaa atcctaactt actcagccca gcatcattct
     1441 tctcctcttg gcagattgtc tgtagccgat tggaggagta caacagccat cagtctttat
     1501 gcaatggaac gcccgaggga cctttacggc gtaatcctgg aaaccatgac aaatccagaa
     1561 ccccaaggct cccctcttca gctgatgtag aattttgcct gagtttgacc caatatgaat
     1621 ctggttccat ggataaagct gccaatttca gctttagaaa tacactggaa ggatttgcta
     1681 gtccacttac tgggatagcg gatgcctctc aaagcagcat gcacaatgcc ttgcacatct
     1741 atatgaatgg aacaatgtcc caggtacagg gatctgccaa cgatcctatc ttccttcttc
     1801 accatgcatt tgttgacagt atttttgagc agtggctccg aaggcaccgt cctcttcaag
     1861 aagtttatcc agaagccaat gcacccattg gacataaccg ggaatcctac atggttcctt
     1921 ttataccact gtacagaaat ggtgatttct ttatttcatc caaagatctg ggctatgact
     1981 atagctatct acaagattca gacccagact cttttcaaga ctacattaag tcctatttgg
     2041 aacaagcgag tcggatctgg tcatggctcc ttggggcggc gatggtaggg gccgtcctca
     2101 ctgccctgct ggcagggctt gtgagcttgc tgtgtcgtca caagagaaag cagcttcctg
     2161 aagaaaagca gccactcctc atggagaaag aggattacca cagcttgtat cagagccatt
     2221 taggatcaac aggatcaaca ggatcaacag gagcagtcga catggcggaa ggatccgtcg
     2281 ccaggcagcc tgacctcttg acctgcgacg atgagccgat ccatatcccc ggtgccatcc
     2341 aaccgcatgg actgctgctc gccctcgccg ccgacatgac gatcgttgcc ggcagcgaca
     2401 accttcccga actcaccgga ctggcgatcg gcgccctgat cggccgctct gcggccgatg
     2461 tcttcgactc ggagacgcac aaccgtctga cgatcgcctt ggccgagccc ggggcggccg
     2521 tcggagcacc gatcactgtc ggcttcacga tgcgaaagga cgcaggcttc atcggctcct
     2581 ggcatcgcca tgatcagctc atcttcctcg agctcgagcc tccccagcgg gacgtcgccg
     2641 agccgcaggc gttcttccgc cgcaccaaca gcgccatccg ccgcctgcag gccgccgaaa
     2701 ccttggaaag cgcctgcgcc gccgcggcgc aagaggtgcg gaagattacc ggcttcgatc
     2761 gggtgatgat ctatcgcttc gcctccgact tcagcggggt ggtgatcgca gaggatcgat
     2821 gcgccgaggt cgagtcaaaa ctaggcctgc actatcctgc ctcagcggtg ccggcgcagg
     2881 cccgtcggct ctataccatc aacccggtac ggatcattcc cgatatcaat tatcggccgg
     2941 tgccggtcac cccagacctc aatccggtca ccgggcggcc gattgatctt agcttcgcca
     3001 tcctgcgcag cgtctcgccc tgccatttgg agttcatgcg caacataggc atgcacggca
     3061 cgatgtcgat ctcgattttg cgcggcgagc gactgtgggg attgatcgtt tgccatcacc
     3121 gaacgccgta ctacgtcgat ctcgatggcc gccaagcctg cgagctagtc gcccaggttc
     3181 tggcctggca gatcggcgtg atggaagagt gagcggccgc gactctagat cataatcagc
     3241 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3301 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3361 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3421 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     3481 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3541 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3601 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3661 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3721 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3781 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3841 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3901 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4021 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4081 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4141 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4201 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4261 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4321 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4381 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4441 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4501 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4561 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4621 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4681 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4741 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4801 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4861 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4921 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4981 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5041 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5101 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5161 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5221 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5281 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5341 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5401 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5461 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5521 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5581 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5641 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6541 atgcat
//
