LOCUS       sequence-335780-        4776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-DsRed2 targeted to the ER lumen, but the encoded 
            protein escapes from the ER into tubular ERGICs due to the 
            N-terminal APV tripeptide after SP cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4776)
  AUTHORS   Yan R, Chen K, Wang B, Xu K
  TITLE     SURF4-induced tubular ERGIC selectively expedites ER-to-Golgi 
            transport.
  JOURNAL   Dev Cell. 2022 Feb 28;57(4):512-525.e8. doi: 
            10.1016/j.devcel.2021.12.018. Epub 2022 Jan 19.
  PUBMED    35051356
REFERENCE   2  (bases 1 to 4776)
  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"; doi: 
            "10.1016/j.devcel.2021.12.018"; journalName: "Dev Cell"; date: 
            "2022-02-28- 28"; volume: "57"; issue: "4"; pages: "512-525.e8"
COMMENT     Sequence Label: pAPV-FLAG-DsRed2-ER-5
FEATURES             Location/Qualifiers
     source          1..4776
                     /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"
     CDS             609..632
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             648..1322
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(828..848)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1240..1263
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1504..1625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1541..1560)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1595..1614
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1632..2087)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1769..1790)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1981..2000
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2114..2218
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2220..2577
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2220..2411
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2243..2263)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2428..2563
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2612..3406
                     /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             2612..3406
                     /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(2666..2685)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3276..3295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3594..3613)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3638..3685
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4014..4602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4503..4522
                     /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 gctagcgcca
      541 ccatgctgct atccgtgccg ttgctgctcg gcctcctcgg cctggccgtc gccgcaccgg
      601 tcgccaccga ctacaaagac gatgacgaca aggaaccggt cgccaccatg gcctcctccg
      661 agaacgtcat caccgagttc atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc
      721 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggccac aacaccgtga
      781 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
      841 tccagtacgg ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc
      901 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
      961 cgaccgtgac ccaggactcc tccctgcagg acggctgctt catctacaag gtgaagttca
     1021 tcggcgtgaa cttcccctcc gacggccccg tgatgcagaa gaagaccatg ggctgggagg
     1081 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc
     1141 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga
     1201 agcccgtgca gctgcccggc tactactacg tggacgccaa gctggacatc acctcccaca
     1261 acgaggacta caccatcgtg gagcagtacg agcgcaccga gggccgccac cacctgttcc
     1321 tgtccggaaa ggacgagctg taagaattct gcagtcgacg gtaccgcggg cccgggatcc
     1381 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     1441 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1501 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1621 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1681 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1741 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1801 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1861 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1921 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1981 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2041 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2101 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2161 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     2221 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2341 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2401 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2461 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2521 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     2581 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2641 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2701 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2761 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     2821 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2881 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2941 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3001 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3061 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3121 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3181 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3241 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3301 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3361 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3421 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3541 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     3601 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3661 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3841 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     3901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4141 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4681 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     4741 ggggtcatta gttcatagcc catatatgga gttccg
//
