LOCUS       sequence-209518-        6888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  AUTHORS   Zeng XC, Bhasin S, Wu X, Lee JG, Maffi S, Nichols CJ, Lee KJ, Taylor
            JP, Greene LE, Eisenberg E
  TITLE     Hsp70 dynamics in vivo: effect of heat shock and protein 
            aggregation.
  JOURNAL   J Cell Sci. 2004 Oct 1. 117(Pt 21):4991-5000.
  PUBMED    15367583
REFERENCE   2  (bases 1 to 6888)
  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 Sci.
            2004 Oct 1. 117(Pt 21):4991-5000."
COMMENT     Sequence Label: pEGFP hsp70
FEATURES             Location/Qualifiers
     source          1..6888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(606..608,609..611,612..1322)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(606..608,609..611,612..1322)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(651..672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1259..1280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3669..3790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3706..3725)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3760..3779
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3797..4252)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3934..3955)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4146..4165
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4279..4383
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4385..4742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4385..4576
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4408..4428)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4593..4728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4777..5571
                     /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             4777..5571
                     /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(4831..4850)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5441..5460
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5759..5778)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5803..5850
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6179..6767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6668..6687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1321 agtactcaga tctcgagctc aagcttatgg ccaaagccgc ggcgatcggc atcgacctgg
     1381 gcaccaccta ctcctgcgtg ggggtgttcc aacacggcaa ggtggagatc atcgccaacg
     1441 accagggcaa ccgcaccacc cccagctacg tggccttcac ggacaccgag cggctcatcg
     1501 gggatgcggc caagaaccag gtggcgctga acccgcagaa caccgtgttt gacgcgaagc
     1561 ggctgatcgg ccgcaagttc ggcgacccgg tggtgcagtc ggacatgaag cactggcctt
     1621 tccaggtgat caacgacgga gacaagccca aggtgcaggt gagctacaag ggggagacca
     1681 aggcattcta ccccgaggag atctcgtcca tggtgctgac caagatgaag gagatcgccg
     1741 aggcgtacct gggctacccg gtgaccaacg cggtgatcac cgtgccggcc tacttcaacg
     1801 actcgcagcg ccaggccacc aaggatgcgg gtgtgatcgc ggggctcaac gtgctgcgga
     1861 tcatcaacga gcccacggcc gccgccatcg cctacggcct ggacagaacg ggcaaggggg
     1921 agcgcaacgt gctcatcttt gacctgggcg ggggcacctt cgacgtgtcc atcctgacga
     1981 tcgacgacgg catcttcgag gtgaaggcca cggccgggga cacccacctg ggtggggagg
     2041 actttgacaa caggctggtg aaccacttcg tggaggagtt caagagaaaa cacaagaagg
     2101 acatcagcca gaacaagcga gccgtgaggc ggctgcgcac cgcctgcgag agggccaaga
     2161 ggaccctgtc gtccagcacc caggccagcc tggagatcga ctccctgttt gagggcatcg
     2221 acttctacac gtccatcacc agggcgaggt tcgaggagct gtgctccgac ctgttccgaa
     2281 gcaccctgga gcccgtggag aaggctctgc gcgacgccaa gctggacaag gcccagattc
     2341 acgacctggt cctggtcggg ggctccaccc gcatccccaa ggtgcagaag ctgctgcagg
     2401 acttcttcaa cgggcgcgac ctgaacaaga gcatcaaccc cgacgaggct gtggcctacg
     2461 gggcggcggt gcaggcggcc atcctgatgg gggacaagtc cgagaacgtg caggacctgc
     2521 tgctgctgga cgtggctccc ctgtcgctgg ggctggagac ggccggaggc gtgatgactg
     2581 ccctgatcaa gcgcaactcc accatcccca ccaagcagac gcagatcttc accacctact
     2641 ccgacaacca acccggggtg ctgatccagg tgtacgaggg cgagagggcc atgacgaaag
     2701 acaacaatct gttggggcgc ttcgagctga gcggcatccc tccggccccc aggggcgtgc
     2761 cccagatcga ggtgaccttc gacatcgatg ccaacggcat cctgaacgtc acggccacgg
     2821 acaagagcac cggcaaggcc aacaagatca ccatcaccaa cgacaagggc cgcctgagca
     2881 aggaggagat cgagcgcatg gtgcaggagg cggagaagta caaagcggag gacgaggtgc
     2941 agcgcgagag ggtgtcagcc aagaacgccc tggagtccta cgccttcaac atgaagagcg
     3001 ccgtggagga tgaggggctc aagggcaaga tcagcgaggc cgacaagaag aaggtgctgg
     3061 acaagtgtca agaggtcatc tcgtggctgg acgccaacac cttggccgag aaggacgagt
     3121 ttgagcacaa gaggaaggag ctggagcagg tgtgtaaccc catcatcagc ggactgtacc
     3181 agggtgccgg tggtcccggg cctgggggct tcggggctca gggtcccaag ggagggtctg
     3241 ggtcaggccc caccattgag gaggtagatt aggggccttt ccaagattgc tgtttttgtt
     3301 ttggagcttc aagactttgc atttcctagt atttctgttt gtcagttctc aatttcctgt
     3361 gtttgcaatg ttgaaatttt ttggtgaagt cggatccacc ggatctagat aactgatcat
     3421 aatcagccat accacatttg tagaggaagg gcgaattcca gcacactggc ggccgttact
     3481 agtggatccg agctcggtac caagcttcga attctgcagt cgacggtacc gcgggcccgg
     3541 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     3601 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3661 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3721 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3781 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3841 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3901 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3961 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4021 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4081 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4141 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4201 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4261 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4321 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4381 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4681 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4741 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5281 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5581 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5641 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5701 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5761 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5821 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5881 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5941 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     6001 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     6061 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6121 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6181 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6241 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6301 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6361 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6421 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6481 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6541 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6601 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6661 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6721 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6781 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6841 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttat
//
