LOCUS       sequence-163237-        5796 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 5796)
  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 5796)
  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-td-Orange2
FEATURES             Location/Qualifiers
     source          1..5796
                     /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             989..1696
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1736..2443
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2584..2705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2621..2640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2675..2694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2712..3167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2849..2870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3061..3080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3194..3298
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3300..3657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3300..3491
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3323..3343)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3508..3643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3692..4486
                     /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             3692..4486
                     /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(3746..3765)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4356..4375
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4674..4693)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4718..4765
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5094..5682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5583..5602
                     /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 cggtcagcac
      961 tagcggcgga agcggcggga caggtggaat ggtgagcaag ggcgaggaga ataacatggc
     1021 catcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     1081 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggctttcaga ccgctaagct
     1141 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcatttcac
     1201 ctacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tcaagctgtc
     1261 cttccccgag ggcttcaagt gggagcgcgt gatgaactac gaggacggcg gcgtggtgac
     1321 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     1381 caccaacttc ccctccgacg gccccgtgat gcagaagaag accatgggct gggaggcctc
     1441 ctccgagcgg atgtaccccg aggacggtgc cctgaagggc aagatcaaga tgaggctgaa
     1501 gctgaaggac ggcggccact acacctccga ggtcaagacc acctacaagg ccaagaagcc
     1561 cgtgcagctg cccggcgcct acatcgtcga catcaagttg gacatcacct cccacaacga
     1621 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     1681 ggacgagttg tacaagagcg gtacctctac cggcggaagc ggcgggacag gtggaatggt
     1741 gagcaagggc gaggagaata acatggccat catcaaggag ttcatgcgct tcaaggtgcg
     1801 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1861 ctacgagggc tttcagaccg ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1921 ctgggacatc ctgtcccctc atttcaccta cggctccaag gcctacgtga agcaccccgc
     1981 cgacatcccc gactacttca agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2041 gaactacgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     2101 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtgatgca
     2161 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggtgccct
     2221 gaagggcaag atcaagatga ggctgaagct gaaggacggc ggccactaca cctccgaggt
     2281 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca tcgtcgacat
     2341 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     2401 cgagggccgc cactccaccg gcggcatgga cgagttgtac aagagcggag gcacaggtta
     2461 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2521 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2581 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2701 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2761 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2821 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2881 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2941 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3001 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3061 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3121 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3181 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3241 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3301 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3361 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3541 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3601 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3661 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3721 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3781 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3841 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3901 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3961 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4021 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4081 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4141 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4201 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4261 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4321 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4381 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4441 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4501 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4561 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4621 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4681 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4741 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4801 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4861 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4921 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4981 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5041 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5221 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     5281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5461 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5761 cctgattctg tggataaccg tattaccgcc atgcat
//
