LOCUS       sequence-356533-        7681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses (via IRES) streptavidin-KDEL and 
            Progranulin-SBP-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Devireddy S, Ferguson SM
  TITLE     Efficient progranulin exit from the ER requires its interaction with
            prosaposin, a Surf4 cargo.
  JOURNAL   J Cell Biol. 2022 Feb 7;221(2):e202104044. doi: 
            10.1083/jcb.202104044. Epub 2021 Dec 17.
  PUBMED    34919127
REFERENCE   2  (bases 1 to 7681)
  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: "2022-02-7"; volume: "221(2):e202104044. doi"; pages: 
            " 10.1083/jcb.202104044. Epub 2021 Dec 17"
COMMENT     Sequence Label: STR-KDEL_PGRN-SBP-mCherry
FEATURES             Location/Qualifiers
     source          1..7681
                     /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"
     CDS             4332..4442
                     /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             4452..5159
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4594..4612)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4862..4881
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5301..5422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5338..5357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5524..5540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5524..5540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5537..5559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5548..5564
                     /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(5572..5578,5579..5596,5597..5602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5617..5638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5755..5772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5926..6514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6006..6025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6685..7476,7477..7545))
                     /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     7308..7327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7546..7650)
                     /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 ggaccctggt gagctgggtg gccttaacag
     2581 cagggctggt ggctggaacg cggtgcccag atggtcagtt ctgccctgtg gcctgctgcc
     2641 tggaccccgg aggagccagc tacagctgct gccgtcccct tctggacaaa tggcccacaa
     2701 cactgagcag gcatctgggt ggcccctgcc aggttgatgc ccactgctct gccggccact
     2761 cctgcatctt taccgtctca gggacttcca gttgctgccc cttcccagag gccgtggcat
     2821 gcggggatgg ccatcactgc tgcccacggg gcttccactg cagtgcagac gggcgatcct
     2881 gcttccaaag atcaggtaac aactccgtgg gtgccatcca gtgccctgat agtcagttcg
     2941 aatgcccgga cttctccacg tgctgtgtta tggtcgatgg ctcctggggg tgctgcccca
     3001 tgccccaggc ttcctgctgt gaagacaggg tgcactgctg tccgcacggt gccttctgcg
     3061 acctggttca cacccgctgc atcacaccca cgggcaccca ccccctggca aagaagctcc
     3121 ctgcccagag gactaacagg gcagtggcct tgtccagctc ggtcatgtgt ccggacgcac
     3181 ggtcccggtg ccctgatggt tctacctgct gtgagctgcc cagtgggaag tatggctgct
     3241 gcccaatgcc caacgccacc tgctgctccg atcacctgca ctgctgcccc caagacactg
     3301 tgtgtgacct gatccagagt aagtgcctct ccaaggagaa cgctaccacg gacctcctca
     3361 ctaagctgcc tgcgcacaca gtgggggatg tgaaatgtga catggaggtg agctgcccag
     3421 atggctatac ctgctgccgt ctacagtcgg gggcctgggg ctgctgccct tttacccagg
     3481 ctgtgtgctg tgaggaccac atacactgct gtcccgcggg gtttacgtgt gacacgcaga
     3541 agggtacctg tgaacagggg ccccaccagg tgccctggat ggagaaggcc ccagctcacc
     3601 tcagcctgcc agacccacaa gccttgaaga gagatgtccc ctgtgataat gtcagcagct
     3661 gtccctcctc cgatacctgc tgccaactca cgtctgggga gtggggctgc tgtccaatcc
     3721 cagaggctgt ctgctgctcg gaccaccagc actgctgccc ccagggctac acgtgtgtag
     3781 ctgaggggca gtgtcagcga ggaagcgaga tcgtggctgg actggagaag atgcctgccc
     3841 gccgggcttc cttatcccac cccagagaca tcggctgtga ccagcacacc agctgcccgg
     3901 tggggcagac ctgctgcccg agcctgggtg ggagctgggc ctgctgccag ttgccccatg
     3961 ctgtgtgctg cgaggatcgc cagcactgct gcccggctgg ctacacctgc aacgtgaagg
     4021 ctcgatcctg cgagaaggaa gtggtctctg cccagcctgc caccttcctg gcccgtagcc
     4081 ctcacgtggg tgtgaaggac gtggagtgtg gggaaggaca cttctgccat gataaccaga
     4141 cctgctgccg agacaaccga cagggctggg cctgctgtcc ctaccgccag ggcgtctgtt
     4201 gtgctgatcg gcgccactgc tgtcctgctg gcttccgctg cgcagccagg ggtaccaagt
     4261 gtttgcgcag ggaggccccg cgctgggacg cccctttgag ggacccagcc ttgagacagc
     4321 tgctggccgt cgacgagaag accactggtt ggcgaggtgg acacgttgtt gaaggactgg
     4381 ctggggaact tgaacaactt cgtgcacgac tggagcatca cccacaaggt caacgtgaac
     4441 cacctgcagg tgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     4501 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     4561 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     4621 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     4681 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     4741 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     4801 tacaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     4861 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     4921 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     4981 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     5041 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     5101 agtacgaacg cgccgagggc cgccactcca ccggtggcat ggacgagctg tacaagtgag
     5161 gcctcgaggg ccaattaatt aactctagat aactgatcat aatcagccat accacatttg
     5221 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     5281 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     5341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5401 ccaaactcat caatgtatct taacgcgtcg agtgcattct agttgtggtt tgtccaaact
     5461 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5581 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6361 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7681 c
//
