LOCUS       sequence-339201-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Tumor Protein D52 like 2 (TPD54/TPD52L2) with 
            N-terminal GFP-FKBP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5576)
  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 5576)
  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: GFP-FKBP-TPD54
FEATURES             Location/Qualifiers
     source          1..5576
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1326)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(613..615,616..618,619..1326)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(1375..1377,1378..1698)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    2364..2485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2401..2420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2455..2474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2492..2947)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2629..2650)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2841..2860
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2974..3078
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3080..3437
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3080..3271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3103..3123)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3288..3423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3472..4266
                     /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             3472..4266
                     /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(3526..3545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4136..4155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4454..4473)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4498..4545
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4874..5462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5363..5382
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtaccgaa tcgatgtgaa tgggggccct gaacccgcca agaacctgta caccatggga
     1381 gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc
     1441 tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac
     1501 agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa
     1561 ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc
     1621 tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg
     1681 gagcttctaa aactggaacg gtacaagtcc ggactcagat ctcgagacat ggactccgcc
     1741 ggccaagata tcaacctgaa ttctcctaac aaaggtctgc tgtctgactc catgacggat
     1801 gttcctgtcg acacaggtgt ggctgcccgg actcctgctg ttgagggtct gacagaggct
     1861 gaggaggagg agctcagggc tgagcttacc aaggtggaag aggaaattgt cactctgcgc
     1921 caggtcctgg cagccaagga gaggcactgt ggagagctca agaggaggct gggcctctcc
     1981 accctggggg agctgaaaca gaacctgtcc aggagctggc atgacgtgca ggtctctagc
     2041 gcctatgtga aaacttctga gaaacttgga gagtggaatg agaaagtgac ccagtcagac
     2101 ctctacaaga agactcagga aactctttca caggcaggac agaagacttc agctgccctg
     2161 tccacagtgg gctctgccat cagcaggaag cttggagaca tgaggaactc tgcgaccttc
     2221 aagtcgtttg aggaccgagt tgggaccata aagtctaagg ttgtgggtga cagagagaac
     2281 ggcagtgaca acctcccttc ctcagcgggg agtggtgaca agcccctgtc ggatcccgca
     2341 cctttctaac aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2401 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2461 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2521 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2581 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2641 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2701 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2761 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2821 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2881 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2941 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3001 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3061 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3121 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3181 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3241 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3301 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3361 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3421 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3481 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3541 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3601 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3661 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3721 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3781 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3841 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3901 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3961 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4021 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4081 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4141 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4201 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4261 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4441 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4501 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4561 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4621 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4681 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4741 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4801 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4861 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4921 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4981 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5041 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5101 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5161 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5221 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5281 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5341 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5401 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5461 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5521 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
