LOCUS       sequence-487700-        5453 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Mammalian secretory granule marker using BDNF mCherry marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 5453)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: BDNF mCherry
FEATURES             Location/Qualifiers
     source          1..5453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1342..1351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1348..2058
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1493..1511)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1761..1780
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2181..2302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2218..2237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2272..2291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2309..2764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2446..2467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2658..2677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2791..2895
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2897..3254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2897..3088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2920..2940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3105..3240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3289..4083
                     /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             3289..4083
                     /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(3343..3362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3953..3972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4271..4290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4315..4362
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4691..5279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5180..5199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcgagtga tgaccatcct tttccttact
      601 atggttattt catacttcgg ttgcatgaag gctgcgccca tgaaagaagc aaacgtccac
      661 ggacaaggca acttggccta cccagctgtg cggacccatg ggactctgga gagcgtgaat
      721 gggcccaggg caggttcgag aggtctgacg acgacgtccc tggctgacac ttttgagcac
      781 gtgatcgaag agctgctgga tgaggaccag aaggttcggc ccaacgaaga aaaccataag
      841 gacgcggact tgtacacttc ccgggtgatg ctcagcagtc aagtgccttt ggagcctcct
      901 ctgctctttc tgctggagga atacaaaaat tacctggatg ccgcaaacat gtctatgagg
      961 gttcggcgcc actccgaccc cgcccgccgt ggggagctga gcgtgtgtga cagtattagc
     1021 gagtgggtca cagcggcaga taaaaagact gcagtggaca tgtccggtgg gacggtcaca
     1081 gtcctggaga aagtcccggt atcaaaaggc caactgaagc aatatttcta cgagaccaag
     1141 tgtaatccca tgggttacac gaaggaaggc tgcaggggca tagacaaaag gcactggaac
     1201 tcgcaatgcc gaactaccca atcgtatgtt cgggccctta ctatggatag caaaaagaga
     1261 attggctggc ggttcataag gatagacact tcctgtgtat gtacactgac cattaaaagg
     1321 ggaagaatgg atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
     1381 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1441 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1501 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     1561 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     1621 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     1681 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1741 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1801 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1861 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1921 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1981 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2041 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     2101 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2161 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2221 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2281 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     2341 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2401 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2461 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2521 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2581 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2641 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2701 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2761 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2821 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2881 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3001 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3061 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3181 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3241 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3301 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3361 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3421 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     3481 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3541 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3601 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3661 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3721 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3781 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     3841 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3901 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3961 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4021 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4081 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4141 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4201 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4261 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4321 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4381 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4441 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4501 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     4561 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4621 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4681 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4741 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4801 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4861 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4921 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4981 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5041 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5101 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5161 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5221 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5281 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5341 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     5401 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccg
//
