LOCUS       sequence-338995-        6331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of untagged Tumor Protein D52 like 2 (TPD54/TPD52L2) and 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6331)
  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 6331)
  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-TPD54
FEATURES             Location/Qualifiers
     source          1..6331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1285..1859,1860..1862,1863..1871)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1452..1469)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1679..1698
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1872..1874,1875..1877,1878..2588)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1872..1874,1875..1877,1878..2588)
                     /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(1917..1938)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2178..2197)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2525..2546
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2619..3215
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2619..2638)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3115..3135
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3347..3468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3384..3403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3438..3457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3482..3501)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3483..3501)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3508..3525)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3508..3524)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3517..3539)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3669..4124)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3806..3827)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4018..4037
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4153..4257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4258..4326,4327..5118)
                     /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(4476..4495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5289..5877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5778..5797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6031..6048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6165..6186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6201..6206,6207..6224,6225..6231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6239..6255
                     /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     6244..6266
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6263..6279
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6263..6279
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6300..6318
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tggatccccc gggctgcagg aattcgatat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      601 gtcagatccg atggactccg ccggccaaga tatcaacctg aattctccta acaaaggtct
      661 gctgtctgac tccatgacgg atgttcctgt cgacacaggt gtggctgccc ggactcctgc
      721 tgttgagggt ctgacagagg ctgaggagga ggagctcagg gctgagctta ccaaggtgga
      781 agaggaaatt gtcactctgc gccaggtcct ggcagccaag gagaggcact gtggagagct
      841 caagaggagg ctgggcctct ccaccctggg ggagctgaaa cagaacctgt ccaggagctg
      901 gcatgacgtg caggtctcta gcgcctatgt gaaaacttct gagaaacttg gagagtggaa
      961 tgagaaagtg acccagtcag acctctacaa gaagactcag gaaactcttt cacaggcagg
     1021 acagaagact tcagctgccc tgtccacagt gggctctgcc atcagcagga agcttggaga
     1081 catgaggaac tctgcgacct tcaagtcgtt tgaggaccga gttgggacca taaagtctaa
     1141 ggttgtgggt gacagagaga acggcagtga caacctccct tcctcagcgg ggagtggtga
     1201 caagcccctg tcggatcccg cacctttcta agctcaagct tcgaattctg cagtcgacgg
     1261 taccgcgggc ccgggatcga tccgcccctc tccctccccc ccccctaacg ttactggccg
     1321 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     1381 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     1441 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     1501 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     1561 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     1621 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     1681 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     1741 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     1801 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     1861 tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1921 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1981 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2041 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2101 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2161 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2221 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2281 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2341 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2401 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2461 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2521 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2581 tgtacaagtc cggactcaga tctaggagac gaccttccat gaccgagtac aagcccacgg
     2641 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     2701 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     2761 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     2821 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     2881 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     2941 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     3001 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     3061 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     3121 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgagtgc ccgaaggacc
     3181 gcgcgacctg gtgcatgacc cgcaagcccg gtgcccggga tccaccggat ctagataact
     3241 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3301 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaaa tcaccaattc
     3481 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     3541 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3601 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3661 atggcgcgaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3721 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3781 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3841 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3901 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3961 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4021 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4081 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccagg tggcactttt
     4141 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4201 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4261 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4321 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4381 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4441 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4501 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4561 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4621 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4681 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4741 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4801 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4861 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4921 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4981 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5041 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5101 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5161 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5221 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5281 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5341 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5401 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5461 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5521 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5581 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5641 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5701 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5761 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5821 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5941 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6001 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6061 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6121 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6181 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6241 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgga
     6301 attaaccctc actaaaggga acaaaagctg g
//
