LOCUS       sequence-413648-        5039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Bet1-mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5039)
  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 5039)
  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: Bet1-mCherry
FEATURES             Location/Qualifiers
     source          1..5039
                     /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      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             994..1704
                     /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(1139..1157)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1407..1426
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1827..1948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1864..1883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1918..1937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1955..2410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2092..2113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2304..2323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2437..2541
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2543..2900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2543..2734
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2566..2586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2751..2886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2935..3729
                     /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             2935..3729
                     /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(2989..3008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3599..3618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3917..3936)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3961..4008
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4337..4925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4826..4845
                     /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 atctcgagat gcgccgcgcc ggcctgggcg agggcgtgcc ccccggcaac
      661 tacggcaact acggctacgc caacagcggc tacagcgcct gcgaggagga gaacgagcgc
      721 ctgaccgaga gcctgcgcag caaggtgacc gccatcaaga gcctgagcat cgagatcggc
      781 cacgaggtga agacccagaa caagctgctg gccgagatgg acagccagtt cgacagcacc
      841 accggcttcc tgggcaagac catgggcaag ctgaagatcc tgagccgcgg cagccagacc
      901 aagctgctgt gctacatgat gctgttcagc ctgttcgtgt tcttcatcat ctactggatc
      961 atcaagctgc gcatggatcc accggtcgcc accatggtga gcaagggcga ggaggataac
     1021 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1081 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1141 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     1201 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     1261 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     1321 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     1381 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1441 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     1501 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     1561 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     1621 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     1681 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     1741 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1801 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1861 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1921 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     1981 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2041 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2101 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2161 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2221 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2281 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2341 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2401 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2461 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2521 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2581 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2641 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2761 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2821 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2881 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2941 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3001 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3061 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3121 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3181 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3241 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3301 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3361 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3421 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3481 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3541 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3601 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3661 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3721 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3781 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3841 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3901 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3961 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4021 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4081 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4141 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4201 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4261 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4321 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4381 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4441 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4501 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4561 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4621 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4681 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4741 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4801 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4861 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4921 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4981 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
