LOCUS       sequence-290575-        6439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Climp63.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6439)
  AUTHORS   Shibata Y, Voss C, Rist JM, Hu J, Rapoport TA, Prinz WA, Voeltz GK
  TITLE     The reticulon and DP1/Yop1p proteins form immobile oligomers in the 
            tubular endoplasmic reticulum.
  JOURNAL   J Biol Chem. 2008 Jul 4. 283(27):18892-904.
  PUBMED    18442980
REFERENCE   2  (bases 1 to 6439)
  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 Biol 
            Chem. 2008 Jul 4. 283(27):18892-904."
COMMENT     Sequence Label: mCh-Climp63
FEATURES             Location/Qualifiers
     source          1..6439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1327
                     /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(765..783)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1033..1052
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3234..3355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3325..3344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3362..3817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3711..3730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3844..3948
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3950..4307
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3950..4141
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3973..3993)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4158..4293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4342..5136
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4342..5136
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4396..4415)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5006..5025
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5324..5343)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5368..5415
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5744..6332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6233..6252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
      661 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
      721 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      781 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
      841 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
      901 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      961 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1021 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1081 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     1141 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1201 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     1261 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1321 gtacaagtcc ggactcagat ctcgagctcc ctcggccaaa caaaggggct ccaagggcgg
     1381 ccacggcgcc gcgagcccct cggacaaggg cgcccacccg tcgggcggcg cggatgacgt
     1441 ggcgaagaag ccgccggcgg cgccgcagca gccgcagcct cccgcgccgc acccgccgca
     1501 gcacccgcag aaccaggcgc accggggcgg ccaccgcggc cggtcctcag ccgccaccgc
     1561 caacgcgtcc tccgcgtcct gctcgcgcag gcttggccgg gtgctcaact ttctcttcta
     1621 cctctccctg gtggcggcgg ccgccttctc gggctggtat gtccatcacg tcctggagga
     1681 ggtccagcag gtccggcgtg gccaccagga cttctcccgt cagagggatg agctgggcca
     1741 gggcttgcag ggcgtcgagc agaaggtcca atccctgcaa gccacatttg ggactttcga
     1801 gtccctcctg agaaactccc agcataaaca ggatctcaca gagaaagccg tgaaggaagg
     1861 ggagagcgag ctcaaccgaa ttagcgaagt tctacagaag ttgcagaatg agattctcaa
     1921 agatctgtcg gacgggatcc acgtggtgaa ggatgcccgg gagcgggact tcacgtccct
     1981 ggagaacaca gtcgaggagc gcctgacgga gcttaccaag tctatcaatg acaacatcgc
     2041 catcttcact gatgtccaga agaggagcca gaaggagatc aatgaggtca agatgaaggt
     2101 ggcctccctg gaagaatcca agggggatcg cagtcaggac gtgaaaactc tgaaggacgc
     2161 cgtgaaggag gtccaggcct ccatgatgtc gagggagagg gacattgagg ccctgaagag
     2221 ttcccttcag acgatggagt ccgacgtcta cacagaggtg cgggaactgg tgagcctcaa
     2281 gcaggagcaa caggcgttca agcaggcggc cgactcagag cgcctggctc tgcaggccct
     2341 cacggagaag cttctgcggt ccgaggagtc ctcctctcgt ctcccagagg acatccggag
     2401 actggaggaa gagctacagc agctgaaggt cggggcccat gggtcagagg aaggtgccgt
     2461 cttcaaagac tccaaagcct tagaggaact gcagaggcag attgagggcc tgggtgccag
     2521 gctccagtat gtagaggacg gggtctactc aatgcaggtg gcttctgcac gccacaccga
     2581 aagcctggag tcgctcctgt ccaagagcca ggagtacgag caacgcctgg ctatgctgca
     2641 ggagcacgtg ggaaacctgg gctcctcatc tgacctggcc agcacagtca ggagcctggg
     2701 ggagacccag ctggcgctgt ccagcgactt gaaggagctg aagcagagcc tgggtgagct
     2761 gcccggtacc gtggagtcac tgcaggagca agtgctctca ctgctcagcc aggaccaagc
     2821 ccaggccgag ggcttgcctc ctcaggactt cctggacaga ctgtcctctc tagacaacct
     2881 gaaatcctca gtgagccaag tggagtcaga cttgaaaatg ctcaggactg ccgtggacag
     2941 tttggtggcc tattccgtca aaatcgaaac gaatgagaat aacttagagt cagccaaggg
     3001 cttgctggat gacctgcgga acgacctgga taggttgttt ctgaaagttg agaagatcca
     3061 tgaaaagatc taagaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     3121 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3181 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     3361 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3421 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3481 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3541 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3601 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3661 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3721 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3781 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3841 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3901 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3961 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4021 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4141 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4201 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4261 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4321 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4501 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4861 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5101 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5161 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5221 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5281 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5341 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5401 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5461 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5521 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5581 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5641 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5701 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5761 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5821 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5881 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5941 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6001 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6061 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6121 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6361 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6421 tggataaccg tattaccgc
//
