LOCUS       sequence-335798-        5936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ER-targeted streptavidin-KDEL (hook) and SBP-Dendra2 
            targeted to the ER lumen with an N-terminal APV tripeptide (cargo) 
            for RUSH..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5936)
  AUTHORS   Yan R, Chen K, Wang B, Xu K
  TITLE     SURF4-induced tubular ERGIC selectively expedites ER-to-Golgi 
            transport.
  JOURNAL   Dev Cell. 2022 Feb 28;57(4):512-525.e8. doi: 
            10.1016/j.devcel.2021.12.018. Epub 2022 Jan 19.
  PUBMED    35051356
REFERENCE   2  (bases 1 to 5936)
  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: 
            "10.1016/j.devcel.2021.12.018"; journalName: "Dev Cell"; date: 
            "2022-02-28- 28"; volume: "57"; issue: "4"; pages: "512-525.e8"
COMMENT     Sequence Label: pStr-KDEL_APV-SBP-Dendra2-ER-5
FEATURES             Location/Qualifiers
     source          1..5936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1412..1641
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1624..1643
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1713..2287,2288..2290,2291..2299)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1880..1897)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2107..2126
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2373..2483
                     /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             2499..3188
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     polyA_signal    3322..3443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3359..3378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3545..3561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3545..3561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3558..3580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3569..3585
                     /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(3593..3599,3600..3617,3618..3623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3638..3659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3776..3793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3947..4535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4027..4046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4706..5497,5498..5566))
                     /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     5329..5348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5567..5671)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5746..5765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcgat atctgcggcc tagctagcgc ttaaggcctg ttaaccggta
      721 tggatgtatg cgtccgtctt gccctgtggc tcctctgggg actcctcctg caccagggcc
      781 agagcctcag ccatagtcac agtgagaagg cgacaggaac cagctcgggc gccggtttaa
      841 acgaccctag caaagactca aaagctcagg tgtccgctgc cgaggctggc attactggaa
      901 catggtacaa tcagctcggg agcaccttta ttgtgactgc tggagccgat ggagccctca
      961 ccggaacata cgaatctgct gtgggaaacg ccgaatcacg gtacgtcctc actggccgat
     1021 acgatagtgc ccctgccacc gacggatctg ggactgccct gggatggact gtcgcttgga
     1081 aaaacaacta ccggaatgct cattctgcca caacatggag tggacagtac gtgggaggcg
     1141 ctgaggctag aatcaataca cagtggctgc tcacatctgg cacaaccgag gcaaatgctt
     1201 ggaaatccac cctggtggga catgacacat tcaccaaagt gaaaccctcc gccgcttcaa
     1261 tcgatgccgc caaaaaagcc ggagtcaaca acggcaatcc tctggatgcc gtccagcagc
     1321 tcggaccgaa agatgaactg tgagcggccg catagataac tgatccagtg tgctggaatt
     1381 aattcgctgt ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg
     1441 acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc
     1501 gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag aaaagacaat ctttttgttg
     1561 tcaagcttga ggtgtggcag gcttgagatc tggccataca cttgagtgac aatgacatcc
     1621 actttgcctt tctctccaca ggtgtccact cccaggtcca actgcaggtc gagcatgcat
     1681 ctagggcggc caattccgcc cctctccccc cccccctctc cctccccccc ccctaacgtt
     1741 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     1801 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     1861 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     1921 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     1981 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2041 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2101 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2161 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     2221 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     2281 tgataagctt gccacaaccc gggaggatgc tgctatccgt gccgttgctg ctcggcctcc
     2341 tcggcctggc cgtcgccgca ccggtcgcca ccgacgagaa gaccactggt tggcgaggtg
     2401 gacacgttgt tgaaggactg gctggggaac ttgaacaact tcgtgcacga ctggagcatc
     2461 acccacaagg tcaacgtgaa ccagcaccgg gcgccaccat gaacaccccg ggaattaacc
     2521 tgatcaagga ggacatgcgc gtgaaggtgc acatggaggg caacgtgaac ggccacgcct
     2581 tcgtgatcga gggcgagggc aagggcaagc cctacgaggg cacccagacc gccaacctga
     2641 ccgtgaagga gggcgccccc ctgcccttca gctacgacat cctgaccacc gccgtgcact
     2701 acggcaaccg ggtgttcacc aagtaccccg aggacatccc cgactacttc aagcagagct
     2761 tccccgaggg ctacagctgg gagcgcacca tgaccttcga ggacaagggc atctgcacca
     2821 tccgcagcga catcagcctg gagggcgact gcttcttcca gaacgtgcgc ttcaagggca
     2881 ccaacttccc ccccaacggc cccgtgatgc agaagaagac cctgaagtgg gagcccagca
     2941 ccgagaagct gcacgtgcgc gacggcctgc tggtgggcaa catcaacatg gccctgctgc
     3001 tggagggcgg cggccactac ctgtgcgact tcaagaccac ctacaaggcc aagaaggtgg
     3061 tgcagctgcc cgacgcccac ttcgtggacc accgcatcga gatcctgggc aacgacagcg
     3121 actacaacaa ggtgaagctg tacgagcacg ccgtggcccg ctacagcccc ctgcccagcc
     3181 aggtgtggtc cggaaaggac gagctgtaaa taactgatca taatcagcca taccacattt
     3241 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3301 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3421 tccaaactca tcaatgtatc ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac
     3481 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3541 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3601 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3661 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3721 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3781 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4381 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4681 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4741 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4801 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4861 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4921 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4981 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5041 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5101 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5161 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5221 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5281 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5341 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5401 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5461 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5521 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5581 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5641 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5701 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     5761 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     5821 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     5881 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
