LOCUS       sequence-339433-        4546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Tumor Protein D52 (TPD52) with N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4546)
  AUTHORS   Larocque G, Moore DJ, Sittewelle M, Kuey C, Hetmanski JHR, La-Borde 
            PJ, Wilson BJ, Clarke NI, Caswell PT, Royle SJ
  TITLE     Intracellular nanovesicles mediate alpha5beta1 integrin trafficking 
            during cell migration.
  JOURNAL   J Cell Biol. 2021 Oct 4;220(10). pii: 212493. doi: 
            10.1083/jcb.202009028. Epub 2021 Jul 21.
  PUBMED    34287617
REFERENCE   2  (bases 1 to 4546)
  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: "2021-10-4"; volume: "220(10). pii: 212493. doi"; 
            pages: " 10.1083/jcb.202009028. Epub 2021 Jul 21"
COMMENT     Sequence Label: FLAG-TPD52
FEATURES             Location/Qualifiers
     source          1..4546
                     /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    1334..1455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1371..1390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1425..1444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1462..1917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1599..1620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1811..1830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1944..2048
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2050..2407
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2050..2241
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2073..2093)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2258..2393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2442..3236
                     /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             2442..3236
                     /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(2496..2515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3106..3125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3424..3443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3468..3515
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3844..4432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4333..4352
                     /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 atctatggat tgtagagaga
      661 tggacttata tgaggactac cagtccccgt ttgattttga tgcaggagtg aacaaaagct
      721 atctctactt gtctcctagt ggaaattcat ctccacccgg atcacctact cttcagaaat
      781 ttggtctgct gagaacagac ccagtccctg aggaaggaga agatgttgct gccacgatca
      841 gtgccacaga gaccctctcg gaagaggagc aggaagagct aagaagagaa cttgcaaagg
      901 tagaagaaga aatccagact ctgtctcaag tgttagcagc aaaagagaag catctagcag
      961 agatcaagcg gaaacttgga atcaattctc tacaggaact aaaacagaac attgccaaag
     1021 ggtggcaaga cgtgacagca acatctgctt acaagaagac atctgaaacc ttatcccagg
     1081 ctggacagaa ggcctcagct gctttttcgt ctgttggctc agtcatcacc aaaaagctgg
     1141 aagatgtaaa aaactcccca acttttaaat catttgaaga aaaggtcgaa aacttaaagt
     1201 ctaaagtagg gggaaccaag cctgctggtg gtgattttgg agaagtcttg aattcggctg
     1261 caaatgctag tgccaccacc acggagcctc ttccagaaaa gacacaggag agcctgtgac
     1321 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1381 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1441 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1501 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1561 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1621 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1681 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1741 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1801 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1861 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     1921 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1981 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2041 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2101 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2161 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2221 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2281 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2341 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2401 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2461 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2521 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2581 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     2641 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2701 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2761 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2821 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     2881 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     2941 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3001 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3061 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3121 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3181 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3241 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3301 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3361 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     3421 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3481 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3541 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3601 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3661 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     3721 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3781 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3841 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3901 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3961 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4021 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4081 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4141 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4201 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4261 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4321 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4381 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4441 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4501 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
