LOCUS       sequence-259745-        5926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an Str-KDEL Hook for the RUSH system and separately 
            Sialyltransferase-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5926)
  AUTHORS   McCaughey J, Stevenson NL, Cross S, Stephens DJ
  TITLE     ER-to-Golgi trafficking of procollagen in the absence of large 
            carriers.
  JOURNAL   J Cell Biol. 2018 Dec 26. pii: jcb.201806035. doi: 
            10.1083/jcb.201806035.
  PUBMED    30587510
REFERENCE   2  (bases 1 to 5926)
  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. 2018 Dec 26. pii: jcb.201806035. doi: 10.1083/jcb.201806035."
COMMENT     Sequence Label: pStr-KDEL-IRES-ST-mCherry
FEATURES             Location/Qualifiers
     source          1..5926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(212..231)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          214..443
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1207..1226)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1208..1226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1271..1474)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1300..1320)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1475..1854
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2026..2045
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2120..2224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2225..2293,2294..3085)
                     /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(2443..2462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3256..3844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3745..3764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3998..4015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4132..4153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4168..4173,4174..4191,4192..4198)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4206..4222
                     /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     4211..4233
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4230..4246
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4230..4246
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4348..4469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4413..4432
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4623..5330)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4898..4917)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5167..5185
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    join(5482..5490,5491..5493,5494..5926,1..142)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5655..5674)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5884..5901
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 caagaagaca gggccaggtt tccgggccct cacattgcca aaagacggca atatggtgga
       61 aaataacata tagacaaacg cacaccggcc ttattccaag cggcttcggc cagtaacgtt
      121 aggggggggg gagggagagg ggggggggga gaggggcgga attggccgcc ctagatgcat
      181 gctcgacctg cagttggacc tgggagtgga cacctgtgga gagaaaggca aagtggatgt
      241 cattgtcact caagtgtatg gccagatctc aagcctgcca cacctcaagc ttgacaacaa
      301 aaagattgtc ttttctgacc agatggacgc ggccaccctc aaaggcatca ccgcgggcca
      361 ggtgaatatc aaatcctcct cgtttttgga aactgacaat cttagcgcag aagtcatgcc
      421 cgcttttgag agggagtact caccccaaca gctggccctc gcagacagcg aattaattcc
      481 agcacactgg atcagttatc tatgcggccg ctcacagttc atctttcggt ccgagctgct
      541 ggacggcatc cagaggattg ccgttgttga ctccggcttt tttggcggca tcgattgaag
      601 cggcggaggg tttcactttg gtgaatgtgt catgtcccac cagggtggat ttccaagcat
      661 ttgcctcggt tgtgccagat gtgagcagcc actgtgtatt gattctagcc tcagcgcctc
      721 ccacgtactg tccactccat gttgtggcag aatgagcatt ccggtagttg tttttccaag
      781 cgacagtcca tcccagggca gtcccagatc cgtcggtggc aggggcacta tcgtatcggc
      841 cagtgaggac gtaccgtgat tcggcgtttc ccacagcaga ttcgtatgtt ccggtgaggg
      901 ctccatcggc tccagcagtc acaataaagg tgctcccgag ctgattgtac catgttccag
      961 taatgccagc ctcggcagcg gacacctgag cttttgagtc tttgctaggg tcgtttaaac
     1021 cggcgcccga gctggttcct gtcgccttct cactgtgact atggctgagg ctctggccct
     1081 ggtgcaggag gagtccccag aggagccaca gggcaagacg gacgcataca tccataccgg
     1141 ttaacaggcc ttaagcgcta gctaggccgc agatatcgat ccgagctcgg taccaagctt
     1201 gggtctccct atagtgagtc gtattaattt cgataagcca gtaagcagtg ggttctctag
     1261 ttagccagag agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     1321 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     1381 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     1441 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     1501 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     1561 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     1621 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     1681 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     1741 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     1801 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcaacgcg
     1861 tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca
     1921 tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag
     1981 cgacctccaa cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga
     2041 ttgtactgag agtgcaccat aggggatcgg gagatctccc gatccgtcga cgtcaggtgg
     2101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3361 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4021 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4081 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4141 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4201 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4261 gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa
     4321 accacaacta gaatgcactc gacgcgttaa gatacattga tgagtttgga caaaccacaa
     4381 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4441 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4501 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     4561 tggctgatta tgatcagtta tctagagtta attaattggc cctcgaggcc ttaaggccgg
     4621 cccttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt
     4681 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag
     4741 gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag
     4801 tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg
     4861 gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg
     4921 tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgtagg
     4981 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac
     5041 ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg
     5101 taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg
     5161 ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
     5221 tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc
     5281 atgaactcct tgatgatggc catgttatcc tcctcgccct tgctcaccat acctgcaggc
     5341 ttggtttgca atttaaagga atcatagtaa ctccctttct tcttttcctt ccacacacag
     5401 atgactgcaa acagaagaaa gaccaggacg cagcagctga actttttctt caggttggtg
     5461 tgaatcatgg cgcgcctccc gggttgtggc aagcttatca tcgtgttttt caaaggaaaa
     5521 ccacgtcccc gtggttcggg gggcctagac gtttttttaa cctcgactaa acacatgtaa
     5581 agcatgtgca ccgaggcccc agatcagatc ccatacaatg gggtaccttc tgggcatcct
     5641 tcagcccctt gttgaatacg cttgaggaga gccatttgac tctttccaca actatccaac
     5701 tcacaacgtg gcactggggt tgtgccgcct ttgcaggtgt atcttataca cgtggctttt
     5761 ggccgcagag gcacctgtcg ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag
     5821 acgttgtttg tcttcaagaa gcttccagag gaactgcttc cttcacgaca ttcaacagac
     5881 cttgcattcc tttggcgaga ggggaaagac ccctaggaat gctcgt
//
