LOCUS       sequence-413646-        5766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Stx5L-mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5766)
  AUTHORS   Linders PTA, Gerretsen ECF, Ashikov A, Vals MA, de Boer R, Revelo 
            NH, Arts R, Baerenfaenger M, Zijlstra F, Huijben K, Raymond K, Muru 
            K, Fjodorova O, Pajusalu S, Ounap K, Ter Beest M, Lefeber D, van den
            Bogaart G
  TITLE     Congenital disorder of glycosylation caused by starting 
            site-specific variant in syntaxin-5.
  JOURNAL   Nat Commun. 2021 Oct 28;12(1):6227. doi: 10.1038/s41467-021-26534-y.
  PUBMED    34711829
REFERENCE   2  (bases 1 to 5766)
  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: "Nat 
            Commun."; date: "2021-10-28"; volume: "12(1):6227. doi"; pages: " 
            10.1038/s41467-021-26534-y"
COMMENT     Sequence Label: Stx5L-mCherry
FEATURES             Location/Qualifiers
     source          1..5766
                     /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      1715..1724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1721..2431
                     /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(1866..1884)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2134..2153
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2554..2675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2591..2610)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2645..2664
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2682..3137)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2819..2840)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3031..3050
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3164..3268
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3270..3627
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3270..3461
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3293..3313)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3478..3613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3662..4456
                     /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             3662..4456
                     /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(3716..3735)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4326..4345
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4644..4663)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4688..4735
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5064..5652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5553..5572
                     /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 attcatgatc ccccgcaagc gctacggcag
      661 caagaacacc gaccagggcg tgtacctggg cctgagcaag acccaggtgc tgagccccgc
      721 caccgccggc agcagcagca gcgacatcgc ccccctgccc ccccccgtga ccctggtgcc
      781 cccccccccc gacaccatga gctgccgcga ccgcacccag gagttcctga gcgcctgcaa
      841 gagcctgcag acccgccaga acggcatcca gaccaacaag cccgccctgc gcgccgtgcg
      901 ccagcgcagc gagttcaccc tgatggccaa gcgcatcggc aaggacctga gcaacacctt
      961 cgccaagctg gagaagctga ccatcctggc caagcgcaag agcctgttcg acgacaaggc
     1021 cgtggagatc gaggagctga cctacatcat caagcaggac atcaacagcc tgaacaagca
     1081 gatcgcccag ctgcaggact tcgtgcgcgc caagggcagc cagagcggcc gccacctgca
     1141 gacccacagc aacaccatcg tggtgagcct gcagagcaag ctggccagca tgagcaacga
     1201 cttcaagagc gtgctggagg tgcgcaccga gaacctgaag cagcagcgca gccgccgcga
     1261 gcagttcagc cgcgcccccg tgagcgccct gcccctggcc cccaaccacc tgggcggcgg
     1321 cgccgtggtg ctgggcgccg agagccacgc cagcaaggac gtggccatcg acatgatgga
     1381 cagccgcacc agccagcagc tgcagctgat cgacgagcag gacagctaca tccagagccg
     1441 cgccgacacc atgcagaaca tcgagagcac catcgtggag ctgggcagca tcttccagca
     1501 gctggcccac atggtgaagg agcaggagga gaccatccag cgcatcgacg agaacgtgct
     1561 gggcgcccag ctggacgtgg aggccgccca cagcgagatc ctgaagtact tccagagcgt
     1621 gaccagcaac cgctggctga tggtgaagat cttcctgatc ctgatcgtgt tcttcatcat
     1681 cttcgtggtg ttcctggccc gggatccacc ggtcgccacc atggtgagca agggcgagga
     1741 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     1801 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1861 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     1921 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     1981 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2041 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2101 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     2161 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2221 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2281 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2341 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2401 caccggcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
     2461 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2521 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2581 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2641 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2701 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2761 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2821 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2881 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2941 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3001 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3061 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3121 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3181 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3241 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3301 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3361 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3421 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3481 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3541 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3601 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3661 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3721 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3781 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3841 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3901 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3961 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4021 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4081 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4141 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4201 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4261 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4321 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4381 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4441 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4501 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4561 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4621 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4681 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4741 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4801 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4861 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4921 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4981 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5041 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5101 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5161 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5221 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5281 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5341 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5401 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5461 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5521 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5581 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5641 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5701 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5761 atgcat
//
