LOCUS       sequence-335801-        6291 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses ER-targeted streptavidin-KDEL (hook) and CXCL9-mCherry-SBP
            targeted to the ER lumen with an N-terminal EPV tripeptide (cargo) 
            for RUSH..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  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 6291)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_EPV-CXCL9-mCherry-SBP
FEATURES             Location/Qualifiers
     source          1..6291
                     /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             3432..3542
                     /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"
     polyA_signal    3677..3798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3714..3733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3900..3916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3900..3916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3913..3935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3924..3940
                     /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(3948..3954,3955..3972,3973..3978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3993..4014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4131..4148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4302..4890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4382..4401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5061..5852,5853..5921))
                     /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     5684..5703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5922..6026)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6101..6120)
                     /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 gggaggcgcg ccatgaagaa gtctggagtc ctgttcctct
     2341 tggggatcat tctgttggta ctgataggtg tacaaggtga accagtcgta agaaaagggc
     2401 ggtgttcctg catttccacc aatcaaggga cgatccatct ccaatccctt aaagacctga
     2461 agcaatttgc tccaagcccc tcttgtgaga agattgaaat catcgctaca ctgaagaatg
     2521 gagtacaaac atgcctgaac ccggattctg cagacgtcaa agagctcatc aagaaatggg
     2581 agaaacaagt ttctcaaaag aagaaacaaa agaacggcaa gaagcatcag aagaagaaag
     2641 tactgaaggt gcggaaaagc caaaggtctc gccaaaagaa gacaaccgtc gactcaggca
     2701 gtagtggaag catggtcagc aagggagaag aggacaacat ggctatcata aaagagttca
     2761 tgaggtttaa agtccatatg gaaggcagcg tcaatggtca cgagttcgag atagaaggag
     2821 agggcgaggg aaggccatac gaaggtacgc agacagccaa gctgaaagtg accaaaggcg
     2881 gtccccttcc cttcgcatgg gacattctta gtccccagtt tatgtacggg tctaaagcgt
     2941 atgttaaaca ccccgcggac attccggatt acctgaagct gtcttttccg gaaggattta
     3001 agtgggagcg ggtcatgaat tttgaagacg gtggggtcgt gaccgttacc caagattcca
     3061 gcctgcaaga cggagaattt atctacaagg tcaagctcag gggcacaaat ttcccatccg
     3121 atggacctgt gatgcaaaag aaaactatgg gatgggaagc ctcaagcgag agaatgtatc
     3181 cagaggacgg tgcattgaaa ggagaaatta agcaaagact caaactcaaa gatggcgggc
     3241 attacgacgc tgaggtcaag accacctata aggcgaagaa acccgtccag ctgcccggcg
     3301 cctataatgt taacattaaa ctggacatca caagccataa cgaagattat acaatcgtcg
     3361 agcagtacga gcgagcggaa ggacggcata gcactggcgg gatggatgag ctttacaagg
     3421 gatccggaag cgacgagaag accactggtt ggcgaggtgg acacgttgtt gaaggactgg
     3481 ctggggaact tgaacaactt cgtgcacgac tggagcatca cccacaaggt caacgtgaac
     3541 catagggcct cgagggccat ctagataact gatcataatc agccatacca catttgtaga
     3601 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3661 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3721 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3781 actcatcaat gtatcttaac gcgtcgagtg cattctagtt gtggtttgtc caaactcatc
     3841 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3901 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3961 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4021 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4921 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4981 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5041 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5101 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5161 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5221 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5281 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5341 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5401 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5461 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5521 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5581 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5641 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5701 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5761 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5821 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5881 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6061 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     6121 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     6181 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6241 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
