LOCUS       sequence-339420-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Tumor Protein D52 like 2 (TPD54/TPD52L2) with 
            N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4498)
  AUTHORS   Larocque G, La-Borde PJ, Clarke NI, Carter NJ, Royle SJ
  TITLE     Tumor protein D54 defines a new class of intracellular transport 
            vesicles.
  JOURNAL   J Cell Biol. 2020 Jan 6;219(1). pii: jcb.201812044. doi: 
            10.1083/jcb.201812044.
  PUBMED    31672706
REFERENCE   2  (bases 1 to 4498)
  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: "J Cell 
            Biol."; date: "2020-01-6"; volume: "219(1). pii: jcb.201812044. 
            doi"; pages: " 10.1083/jcb.201812044"
COMMENT     Sequence Label: FLAG-TPD54
FEATURES             Location/Qualifiers
     source          1..4498
                     /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      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             615..638
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1286..1407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1323..1342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1377..1396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1414..1869)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1551..1572)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1763..1782
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1896..2000
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2002..2359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2002..2193
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2025..2045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2210..2345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2394..3188
                     /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             2394..3188
                     /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(2448..2467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3058..3077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3376..3395)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3420..3467
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3796..4384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4285..4304
                     /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 ccggtgccac catggactac aaagacgatg acgacaagag atctcgagac atggactccg
      661 ccggccaaga tatcaacctg aattctccta acaaaggtct gctgtctgac tccatgacgg
      721 atgttcctgt cgacacaggt gtggctgccc ggactcctgc tgttgagggt ctgacagagg
      781 ctgaggagga ggagctcagg gctgagctta ccaaggtgga agaggaaatt gtcactctgc
      841 gccaggtcct ggcagccaag gagaggcact gtggagagct caagaggagg ctgggcctct
      901 ccaccctggg ggagctgaaa cagaacctgt ccaggagctg gcatgacgtg caggtctcta
      961 gcgcctatgt gaaaacttct gagaaacttg gagagtggaa tgagaaagtg acccagtcag
     1021 acctctacaa gaagactcag gaaactcttt cacaggcagg acagaagact tcagctgccc
     1081 tgtccacagt gggctctgcc atcagcagga agcttggaga catgaggaac tctgcgacct
     1141 tcaagtcgtt tgaggaccga gttgggacca taaagtctaa ggttgtgggt gacagagaga
     1201 acggcagtga caacctccct tcctcagcgg ggagtggtga caagcccctg tcggatcccg
     1261 cacctttcta acaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1321 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1381 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     1441 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     1501 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     1561 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1621 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     1681 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     1741 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     1801 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     1861 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1981 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2041 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2101 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2161 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2221 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2281 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2341 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     2401 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2461 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2521 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     2581 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2641 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2701 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2761 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2821 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2881 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     2941 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3001 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3061 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3121 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3181 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3241 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3301 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3361 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3421 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     3481 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3541 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3601 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     3661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3841 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4381 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4441 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
