LOCUS       sequence-442330-        7489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synchronize the trafficking of gp135/podocalyxin from the ER..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7489)
  AUTHORS   Fourriere L, Kasri A, Gareil N, Bardin S, Bousquet H, Pereira D, 
            Perez F, Goud B, Boncompain G, Miserey-Lenkei S
  TITLE     RAB6 and microtubules restrict protein secretion to focal adhesions.
  JOURNAL   J Cell Biol. 2019 Jul 1;218(7):2215-2231. doi: 
            10.1083/jcb.201805002. Epub 2019 May 29.
  PUBMED    31142554
REFERENCE   2  (bases 1 to 7489)
  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"; doi: 
            "10.1083/jcb.201805002"; journalName: "J Cell Biol"; date: 
            "2019-07-1- 1"; volume: "218"; issue: "7"; pages: "2215-2231"
COMMENT     Sequence Label: Str-KDEL_SBP-EGFP-gp135
FEATURES             Location/Qualifiers
     source          1..7489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1646..1875
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1858..1877
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1947..2521,2522..2524,2525..2533)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2114..2131)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2341..2360
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     2547..2606
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             2607..2717
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             join(2727..2729,2730..3440)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2727..2729,2730..3440)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2769..2790)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3030..3049)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3377..3398
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5109..5230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5146..5165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5332..5348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5332..5348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5345..5367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5356..5372
                     /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)."
     promoter        complement(join(5380..5386,5387..5404,5405..5410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5425..5446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5563..5580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5734..6322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5814..5833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6493..7284,7285..7353))
                     /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     7116..7135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7354..7458)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
      961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgacc
     1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
     1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
     1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
     1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
     1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
     1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
     1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
     1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
     1561 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
     1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     1921 cggccaattc cgcccctctc cccccccccc tctccctccc ccccccctaa cgttactggc
     1981 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     2041 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     2101 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     2161 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     2221 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     2281 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2341 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2401 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     2461 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2521 gcttgccaca acccgggagg cgcgccatgt acaggatgca actcctgtct tgcattgcac
     2581 taagtcttgc acttgtcacg aattccgacg agaagaccac tggttggcga ggtggacacg
     2641 ttgttgaagg actggctggg gaacttgaac aacttcgtgc acgactggag catcacccac
     2701 aaggtcaacg tgaaccacct gcaggtgtga gcaagggcga ggagctgttc accggggtgg
     2761 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2821 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2881 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2941 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     3001 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     3061 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     3121 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     3181 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     3241 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3301 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     3361 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3421 gcatggacga gctgtacaag ggccggcccc aggaaaagag tccccagccc ggcccaaccc
     3481 ccatggccac ttcaacatct accaggcctg ccccagcctc agccccagcc cccaaaagct
     3541 ccgtggcggc ttcggtcccg gccgagcaga acactacacc catgacgacg aaagccccag
     3601 ccacccagag cccctctgca tcccctggaa gctccgtgga gaactcagcc ccggcccagg
     3661 gctcgaccac aacacagcag agcctgagtg ttactactaa ggctgaagct aaggatgcag
     3721 gtggcgtccc cacggcacac gtcaccggct ctgctcggcc tgtcaccagt ggcagccagg
     3781 tcgcagctca ggacccagct gcatccaaag ctccgagcaa ccacagcatc accacaaagc
     3841 ctctggccac agaggccacg agccaggccc cccgtcagac cacagatgtc ggcaccccgg
     3901 gacccactgc tcctcctgtg accaactcga ccagccctga cctgctgggc catgccaccc
     3961 cgaagccctc cgaggggccc cagctcagtt tccccacggc tgctggcagc ctgggccctg
     4021 tcactggctc tggcaccggc tctggcaccc tctccacacc ccaggggaag cctgcgacac
     4081 tgacgcccgt ggccagctca gcagagactc aagggatgcc cagcccgatg ccacccagcc
     4141 cagcctcccc ctcctcctcc ccctttccct cctctccctc cccctccccc gcactgcagc
     4201 cctcgggccc ctcggcggca ggaacagagg acacaaccgg caggggcccc acaagctcca
     4261 gcactgagtt ggcttcaact gccctgcacg gccccagcac gctttccccc acctcggcag
     4321 tgagagacca gagggtaagc tgcgggcctc cggagagacc aaccgagcag ctgctcatcc
     4381 tgaacctcac cagaagcagc ccctgcacag acagccttcc ggaagacaaa ctggtcaccc
     4441 tgctgtgccg agcggccaag cccaccttca accccgccca ggaccagtgc cacgtgctgc
     4501 tggcgcccat gctggggagc cacgcggtgg tggtcaaaga aatcaccatc aagacgaatc
     4561 tgctgcccac ggccgtgttc gagctgctga aggacagatg ggatgacctg agagaggagg
     4621 gcgtcagcga catgcagctg ggggaccagg ggcccccgga ggagaccgag gaccgcttca
     4681 gcctgccgct catcatcacc atcgtctgca tggcctcctt cctgctgctt gtcgcggccc
     4741 tctacggctg ctgccaccag cgcctctccc ataggaagga ccagcaacga ctcacggagg
     4801 agctgcagac cgtggagaac ggttaccatg acaacccgac cctagaagtg atggagacgt
     4861 cggccgagat gcaggagaag aaggtggtca accttaacgg ggagctgggg gacagctgga
     4921 ttgtcccgct ggacaacctg accaaggacg acctggacga ggaggaggac acgcacctct
     4981 aattaattaa ctctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     5041 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     5101 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5221 atgtatctta acgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5281 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5461 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5521 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5581 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
