LOCUS       sequence-372880-        7033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-Climp63 with partial luminal domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  AUTHORS   Wang B, Zhao Z, Xiong M, Yan R, Xu K
  TITLE     The endoplasmic reticulum adopts two distinct tubule forms.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 May 3;119(18):e2117559119. doi: 
            10.1073/pnas.2117559119. Epub 2022 Apr 26.
  PUBMED    35471903
REFERENCE   2  (bases 1 to 7033)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-05-3"; volume: "119(18):e2117559119. 
            doi"; pages: " 10.1073/pnas.2117559119. Epub 2022 Apr 26"
COMMENT     Sequence Label: mCherry-Climp63(1–301)
FEATURES             Location/Qualifiers
     source          1..7033
                     /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"
     CDS             958..1665
                     /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(1103..1121)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1371..1390
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2630..2854
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2900..3328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2987..3006)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3197..3218
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3337..3357)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3342..3671
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3522..3657
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3584..3603
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3738..4532
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3792..3811)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4402..4421
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4706..4827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4743..4762)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4797..4816
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4876..4892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4876..4892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4889..4911)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4900..4916
                     /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(4924..4930,4931..4948,4949..4954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4969..4990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5107..5124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5278..5866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5358..5377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6037..6828,6829..6897))
                     /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     6660..6679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6898..7002)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcatg
      961 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1021 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1081 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1141 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     1201 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1261 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     1321 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1381 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     1441 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     1501 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     1561 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     1621 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtccgg actcagatct
     1681 cgagctccct cggccaaaca aaggggctcc aagggcggcc acggcgccgc gagcccctcg
     1741 gacaagggcg cccacccgtc gggcggcgcg gatgacgtgg cgaagaagcc gccggcggcg
     1801 ccgcagcagc cgcagcctcc cgcgccgcac ccgccgcagc acccgcagaa ccaggcgcac
     1861 cggggcggcc accgcggccg gtcctcagcc gccaccgcca acgcgtcctc cgcgtcctgc
     1921 tcgcgcaggc ttggccgggt gctcaacttt ctcttctacc tctccctggt ggcggcggcc
     1981 gccttctcgg gctggtatgt ccatcacgtc ctggaggagg tccagcaggt ccggcgtggc
     2041 caccaggact tctcccgtca gagggatgag ctgggccagg gcttgcaggg cgtcgagcag
     2101 aaggtccaat ccctgcaagc cacatttggg actttcgagt ccctcctgag aaactcccag
     2161 cataaacagg atctcacaga gaaagccgtg aaggaagggg agagcgagct caaccgaatt
     2221 agcgaagttc tacagaagtt gcagaatgag attctcaaag atctgtcgga cgggatccac
     2281 gtggtgaagg atgcccggga gcgggacttc acgtccctgg agaacacagt cgaggagcgc
     2341 ctgacggagc ttaccaagtc tatcaatgac aacatcgcca tcttcactga tgtccagaag
     2401 aggagccaga aggagatcaa tgaggtcaag atgaaggtgg cctccctgga agaatccaag
     2461 ggggatcgca gtcaggacgt gaaaactctg aaggacgccg tgaaggaggt ccaggcctcc
     2521 atgatgtcga gggagaggga cattgaggcc ctgaagagtt cccttcagac gatggagtcc
     2581 gacgtctact aatctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     2641 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2701 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2761 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2821 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2881 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2941 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3001 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3061 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3121 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3181 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3241 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3301 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3361 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3421 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3481 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3541 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3601 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3661 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3721 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3781 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3841 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3901 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3961 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4021 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4081 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4141 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4201 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4261 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4321 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4381 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4441 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4501 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4561 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4621 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4681 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4741 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4801 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4861 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4921 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4981 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5041 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5101 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5161 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5221 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5281 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5341 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5401 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5461 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5521 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5581 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5641 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5701 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5761 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6961 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7021 gccacctgac gtc
//
