LOCUS       sequence-163238-        5007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a fluorescent granule marker, neuropeptide-Y, for 
            localization and exocytosis studies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5007)
  AUTHORS   Gandasi NR, Vesto K, Helou M, Yin P, Saras J, Barg S
  TITLE     Survey of Red Fluorescence Proteins as Markers for Secretory Granule
            Exocytosis.
  JOURNAL   PLoS One. 2015 Jun 19;10(6):e0127801. doi: 
            10.1371/journal.pone.0127801. eCollection 2015.
  PUBMED    26091288
REFERENCE   2  (bases 1 to 5007)
  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: "PLoS One.";
            date: "2015-06-19"; volume: "10(6):e0127801. doi"; pages: " 
            10.1371/journal.pone.0127801. eCollection 2015"
COMMENT     Sequence Label: NPY-mApple
FEATURES             Location/Qualifiers
     source          1..5007
                     /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"
     regulatory      956..965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             962..1672
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1375..1394
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1795..1916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1832..1851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1886..1905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1923..2378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2060..2081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2272..2291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2405..2509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2511..2868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2511..2702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2534..2554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2719..2854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2903..3697
                     /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             2903..3697
                     /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(2957..2976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3567..3586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3885..3904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3929..3976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4305..4893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4794..4813
                     /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 caagcttagc caccatgcta ggtaacaagc gactggggct
      661 gtccggactg accctcgccc tgtccctgct cgtgtgcctg ggtgcgctgg ccgaggcgta
      721 cccctccaag ccggacaacc cgggcgagga cgcaccagcg gaggacatgg ccagatacta
      781 ctcggcgctg cgacactaca tcaacctcat caccaggcag agatatggaa aacgatccag
      841 cccagagaca ctgatttcag acctcttgat gagagaaagc acagaaaatg ttcccagaac
      901 tcggcttgaa gaccctgcaa tgtggacggt accgcgggcc cgggatccac cggtcgccac
      961 catggtgagc aagggcgagg agaataacat ggccatcatc aaggagttca tgcgcttcaa
     1021 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1081 ccgcccctac gaggcctttc agaccgctaa gctgaaggtg accaagggtg gccccctgcc
     1141 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggtct acattaagca
     1201 cccagccgac atccccgact acttcaagct gtccttcccc gagggcttca ggtgggagcg
     1261 cgtgatgaac ttcgaggacg gcggcattat tcacgttaac caggactcct ccctgcagga
     1321 cggcgtgttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1381 aatgcagaag aagaccatgg gctgggaggc ctccgaggag cggatgtacc ccgaggacgg
     1441 cgccctgaag agcgagatca agaagaggct gaagctgaag gacggcggcc actacgccgc
     1501 cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt
     1561 cgacatcaag ttggacatcg tgtcccacaa cgaggactac accatcgtgg aacagtacga
     1621 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaagcggccg
     1681 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1741 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1801 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1861 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     1921 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1981 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2041 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2101 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2161 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2221 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2281 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2341 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2521 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2821 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     2881 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2941 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3001 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3061 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3121 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3181 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3241 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3301 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3361 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3421 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3481 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3541 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3601 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3661 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3721 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3841 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     3901 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     3961 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4021 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4081 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4141 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4441 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4981 gtggataacc gtattaccgc catgcat
//
