LOCUS       sequence-233050-        8086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an Str-KDEL Hook for the RUSH system and separately 
            mScarleti-Sec23A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  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 8086)
  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: StrKDEL-IRES_mScarleti_Sec23A
FEATURES             Location/Qualifiers
     source          1..8086
                     /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             2550..3242
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELY"
     primer_bind     complement(2683..2701)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2951..2970
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5706..5827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5743..5762)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5929..5945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5929..5945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5942..5964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5953..5969
                     /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(5977..5983,5984..6001,6002..6007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6022..6043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6160..6177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6331..6919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6411..6430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7090..7881,7882..7950))
                     /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     7713..7732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7951..8055)
                     /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 cgcgcctaaa tggtgagcaa gggcgaggca gtgatcaagg
     2581 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     2641 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2701 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2761 gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     2821 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     2881 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     2941 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     3001 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     3061 gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     3121 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     3181 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     3241 acaacctgca gggaatgaca acctatttgg aattcattca acaaaatgaa gaacgagatg
     3301 gagtccgatt tagttggaat gtttggccat caagtcgact ggaagctaca agaatggttg
     3361 ttcctgtggc agccctgttt acaccactga aagagagacc tgacttacca cctattcaat
     3421 atgaacctgt tctgtgtagt aggaccactt gccgtgcagt tttgaatcct ttatgtcaag
     3481 tggattatcg agcaaaactt tgggcttgca acttttgtta ccaaaggaat cagtttccac
     3541 ctagttatgc tggtatatct gaactgaatc agcctgctga acttttacct cagttttcta
     3601 gcattgaata tgtagttctg cgtggtcctc agatgccttt gatattcctc tatgtggttg
     3661 atacttgcat ggaagatgaa gatttacaag ccctgaaaga atccatgcag atgtcattaa
     3721 gtcttttacc acctacagct ttggttggac ttattacttt tgggagaatg gttcaggttc
     3781 atgaacttgg atgtgaaggc atttcaaaaa gctatgtctt cagaggaaca aaagatttgt
     3841 ctgccaaaca actgcaggaa atgctggggc tctctaaagt accagttact caagcaacac
     3901 gtggtcctca ggtacagcag ccacctcctt ccaacagatt cttacaacca gtacagaaaa
     3961 tagacatgaa tctcacagat cttctgggag aactccagcg agacccttgg cctgtaccac
     4021 agggaaagag acctttgcgt tcctctgggg tggcactttc catagctgta ggactgctgg
     4081 agtgtacttt tcccaacact ggtgctcgta tcatgatgtt cattggtggt cctgctactc
     4141 aggggcctgg aatggtggtt ggagatgagt tgaagacacc tataagatcg tggcatgaca
     4201 ttgacaaaga caatgccaaa tatgttaaaa agggaactaa gcattttgaa gcattggcta
     4261 atcgagctgc tacaactggc catgttattg atatctatgc gtgtgcatta gatcagacag
     4321 gtctcctgga gatgaaatgc tgtcccaacc ttactggagg atacatggta atgggtgatt
     4381 ctttcaatac ttccttattc aaacaaactt ttcaaagagt ctttaccaaa gacatgcatg
     4441 gacagtttaa aatgggcttt ggtggtacgc tagaaataaa gacctcaagg gaaataaaga
     4501 tttcaggagc tattggaccc tgtgtgtcac tcaattctaa aggaccctgt gtgtctgaaa
     4561 atgagatagg aacaggtggc acatgtcagt ggaagatatg tggacttagt cccactacaa
     4621 ccttagccat atattttgag gttgtcaatc agcataatgc tccaattcct caaggagggc
     4681 gtggtgcaat ccagtttgtg actcagtatc agcattcaag tgggcagaga cgcatccgag
     4741 tgaccaccat tgctaggaac tgggcagatg ctcaaactca aatccaaaac attgctgcat
     4801 cttttgacca ggaggcagct gccattctta tggcccggct agcaatatat agagcagaaa
     4861 cagaagaagg tccagatgtg cttaggtggc tggacagaca gctcattcga ctgtgtcaga
     4921 aatttggaga atatcataaa gatgacccaa gttccttcag attttcagaa actttctccc
     4981 tttatccaca gtttatgttt catttaagaa gatcttcttt cctgcaagtt tttaacaata
     5041 gtcctgatga gagttcatat tatcgtcacc attttatgcg tcaagatctg acccagtctc
     5101 taattatgat tcagcctatc ctgtatgcgt attcttttag tggaccacca gagccggttc
     5161 ttcttgatag cagtagcatt cttgcagatc gtattcttct catggacaca ttcttccaga
     5221 ttttgattta tcatggtgag accatagcac agtggcggaa gtcaggatac caggatatgc
     5281 ctgagtatga aaatttccgc caccttctgc aagccccagt ggatgatgca caggaaattc
     5341 ttcactccag atttccaatg ccaagataca ttgacactga acatggaggc agccaggccc
     5401 gtttcctcct ttcaaaagtc aacccttcac agactcataa taatatgtat gcctgggggc
     5461 aggagtctgg agcacctatt cttacagatg atgttagttt acaagtgttt atggatcact
     5521 tgaagaaact tgctgtgtcc agtgctgctt aaggccggcc ttaaggcctc gagggccaat
     5581 taattaactc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     5641 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5701 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5821 tatcttaacg cgtcgagtgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5881 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5941 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6001 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6061 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
