LOCUS       sequence-128872-        6165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express cytoplasmic mature CRT..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6165)
  AUTHORS   Goitea VE, Hallak ME
  TITLE     Calreticulin and Arginylated Calreticulin Have Different 
            Susceptibilities to Proteasomal Degradation.
  JOURNAL   J Biol Chem. 2015 Jun 26;290(26):16403-14. doi: 
            10.1074/jbc.M114.626127. Epub 2015 May 12.
  PUBMED    25969538
REFERENCE   2  (bases 1 to 6165)
  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"; doi: 
            "10.1074/jbc.M114.626127"; journalName: "J Biol Chem"; date: 
            "2015-06-26- 26"; volume: "290"; issue: "26"; pages: "16403-14"
COMMENT     Sequence Label: pEGFP N1 Ub-CRT
FEATURES             Location/Qualifiers
     source          1..6165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             647..874
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     regulatory      2105..2114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2111..2113,2114..2116,2117..2830)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2111..2113,2114..2116,2117..2827)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2156..2177)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2417..2436)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2764..2785
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2953..3074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2990..3009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3044..3063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3081..3536)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3218..3239)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3430..3449
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3563..3667
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3669..4026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3669..3860
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3692..3712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3877..4012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4061..4855
                     /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             4061..4855
                     /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(4115..4134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4725..4744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5043..5062)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5087..5134
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5463..6051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5952..5971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attccccgcc gccaccatgc agatctttgt
      661 gaagaccctc actggcaaaa ccatcaccct tgaggtcgag cccagtgaca ccattgagaa
      721 tgtcaaagcc aaaattcaag acaaggaggg tatcccacct gaccagcagc gtctgatatt
      781 tgccggcaaa cagctggagg atggccgcac tctctcagac tacaacatcc agaaagagtc
      841 caccctgcac ctggtgttgc gcctccgcgg tggagagcct gccgtctact tcaaggagca
      901 gtttctggac ggagacgggt ggacttcccg ctggatcgaa tccaaacaca agtcagattt
      961 tggcaaattc gttctcagtt ccggcaagtt ctacggtgac gaggagaaag ataaaggttt
     1021 gcagacaagc caggatgcac gcttttatgc tctgtcggcc agtttcgagc ctttcagcaa
     1081 caaaggccag acgctggtgg tgcagttcac ggtgaaacat gagcagaaca tcgactgtgg
     1141 gggcggctat gtgaagctgt ttcctaatag tttggaccag acagacatgc acggagactc
     1201 agaatacaac atcatgtttg gtcccgacat ctgtggccct ggcaccaaga aggttcatgt
     1261 catcttcaac tacaagggca agaacgtgct gatcaacaag gacatccgtt gcaaggatga
     1321 tgagtttaca cacctgtaca cactgattgt gcggccagac aacacctatg aggtgaagat
     1381 tgacaacagc caggtggagt ccggctcctt ggaagacgat tgggacttcc tgccacccaa
     1441 gaagataaag gatcctgatg cttcaaaacc ggaagactgg gatgagcggg ccaagatcga
     1501 tgatcccaca gactccaagc ctgaggactg ggacaagccc gagcatatcc ctgaccctga
     1561 tgctaagaag cccgaggact gggatgaaga gatggacgga gagtgggaac ccccagtgat
     1621 tcagaaccct gagtacaagg gtgagtggaa gccccggcag atcgacaacc cagattacaa
     1681 gggcacttgg atccacccag aaattgacaa ccccgagtat tctcccgatc ccagtatcta
     1741 tgcctatgat aactttggcg tgctgggcct ggacctctgg caggtcaagt ctggcaccat
     1801 ctttgacaac ttcctcatca ccaacgatga ggcatacgct gaggagtttg gcaacgagac
     1861 gtggggcgta acaaaggcag cagagaaaca aatgaaggac aaacaggacg aggagcagag
     1921 gcttaaggag gaggaagaag acaagaaacg caaagaggag gaggaggcag aggacaagga
     1981 ggatgatgag gacaaagatg aggatgagga ggatgaggag gacaaggagg aagatgagga
     2041 ggaagatgtc cccggccagg ccaaggacga gctgggggta ccgcgggccc gggatccacc
     2101 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2161 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2221 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2281 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2341 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2401 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2461 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2521 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2581 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2641 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2701 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2761 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2821 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2881 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2941 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3001 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3061 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3121 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3181 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3241 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3301 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3361 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3421 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3481 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3661 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3721 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3781 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3841 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3901 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3961 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4021 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4081 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4141 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4201 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4261 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4321 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4381 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4441 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4501 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4561 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4621 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4681 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4741 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4801 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4861 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4921 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4981 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     5041 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     5101 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5161 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5221 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5281 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5341 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5401 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5461 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5521 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5581 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5641 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5701 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5761 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5821 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5881 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5941 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6001 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6061 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6121 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
