LOCUS       sequence-372866-        6706 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 6706)
  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 6706)
  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(1–192)
FEATURES             Location/Qualifiers
     source          1..6706
                     /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"
     primer_bind     complement(2297..2314)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2303..2527
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2573..3001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2660..2679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2870..2891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3010..3030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3015..3344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3195..3330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3257..3276
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3411..4205
                     /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(3465..3484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4075..4094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4379..4500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4416..4435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4470..4489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4549..4565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4549..4565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4562..4584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4573..4589
                     /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(4597..4603,4604..4621,4622..4627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4642..4663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4780..4797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4951..5539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5031..5050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5710..6501,6502..6570))
                     /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     6333..6352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6571..6675)
                     /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 attaatctag agggcccgtt
     2281 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2341 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2401 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2461 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2521 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2941 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3001 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3361 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4201 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4261 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4321 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4381 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4441 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4501 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5461 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
