LOCUS       sequence-339134-        6407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of untagged Tumor Protein D52 (TPD52) and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  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 6407)
  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: pIRES-EGFP-TPD52
FEATURES             Location/Qualifiers
     source          1..6407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1353..1927,1928..1930,1931..1939)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1520..1537)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1747..1766
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1940..1942,1943..1945,1946..2656)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1940..1942,1943..1945,1946..2656)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1985..2006)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2246..2265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2593..2614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2687..3283
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2687..2706)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3183..3203
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3413..3534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3450..3469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3504..3523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3556..3575)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3557..3575)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3585..3602)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3585..3601)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3594..3616)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3742..4198)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3880..3901)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4092..4111
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4225..4329
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4330..4398,4399..5190)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4548..4567)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5361..5949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5850..5869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6103..6120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6237..6258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6273..6278,6279..6296,6297..6303)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6311..6327
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6316..6338
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6335..6351
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6335..6351
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6370..6390
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6372..6390
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tggatccccc gggctgcagg aattcgatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc cgctagctat ggattgtaga gagatggact tatatgagga ctaccagtcc
      661 ccgtttgatt ttgatgcagg agtgaacaaa agctatctct acttgtctcc tagtggaaat
      721 tcatctccac ccggatcacc tactcttcag aaatttggtc tgctgagaac agacccagtc
      781 cctgaggaag gagaagatgt tgctgccacg atcagtgcca cagagaccct ctcggaagag
      841 gagcaggaag agctaagaag agaacttgca aaggtagaag aagaaatcca gactctgtct
      901 caagtgttag cagcaaaaga gaagcatcta gcagagatca agcggaaact tggaatcaat
      961 tctctacagg aactaaaaca gaacattgcc aaagggtggc aagacgtgac agcaacatct
     1021 gcttacaaga agacatctga aaccttatcc caggctggac agaaggcctc agctgctttt
     1081 tcgtctgttg gctcagtcat caccaaaaag ctggaagatg taaaaaactc cccaactttt
     1141 aaatcatttg aagaaaaggt cgaaaactta aagtctaaag tagggggaac caagcctgct
     1201 ggtggtgatt ttggagaagt cttgaattcg gctgcaaatg ctagtgccac caccacggag
     1261 cctcttccag aaaagacaca ggagagcctg tgacctcgag ctcaagcttc gaattctgca
     1321 gtcgacggta ccgcgggccc gggatcgatc cgcccctctc cctccccccc ccctaacgtt
     1381 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     1441 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     1501 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     1561 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     1621 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     1681 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     1741 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     1801 cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg
     1861 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     1921 tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1981 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2041 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2101 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2161 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2221 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2281 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2341 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2401 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2461 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2521 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2581 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2641 ggacgagctg tacaagtccg gactcagatc taggagacga ccttccatga ccgagtacaa
     2701 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     2761 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     2821 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     2881 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     2941 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3001 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3061 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3121 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3181 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3241 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcgggatcca ccggatctag
     3301 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3361 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcga
     3541 tcaagctcca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     3601 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3661 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3721 cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3781 atttttgtta aatcagctca tttttttaac caataggccg aaatcggcaa aatcccttat
     3841 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3901 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3961 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4021 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4081 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4141 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4201 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4261 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4321 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4381 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4441 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4501 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4561 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4621 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4681 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4741 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4801 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4861 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4921 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4981 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5041 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5101 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5161 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5221 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5281 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5341 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5401 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5461 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5521 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5581 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5641 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5701 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5761 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5821 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5881 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5941 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6001 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6061 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6121 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6181 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6241 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6301 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6361 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggctag
//
