LOCUS       sequence-223356-        6716 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 6716)
  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 6716)
  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 K71E
FEATURES             Location/Qualifiers
     source          1..6716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3444..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3535..3554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3572..4027)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3709..3730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3921..3940
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4054..4158
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4160..4517
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4160..4351
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4183..4203)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4368..4503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4552..5346
                     /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             4552..5346
                     /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(4606..4625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5216..5235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5534..5553)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5578..5625
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5954..6542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6443..6462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt tggccaaagc cgcggcagtc ggcatcgacc tgggcaccac ctactcctgc
     1321 gtgggggtgt tccaacacgg caaggtggag atcatcgcca acgaccaggg caaccgcacc
     1381 acccccagct acgtggcctt cacggacacc gagcggctca tcggggatgc ggccaagaac
     1441 caggtggcgc tgaacccgca gaacaccgtg tttgacgcgg agcgcctgat cggccgtaag
     1501 ttcggcgacc cggtggtgca gtcggacatg aagcactggc ctttccaggt gatcaacgac
     1561 ggagacaagc ccaaggtgca ggtgagctac aagggggaga ccaaggcatt ctaccccgag
     1621 gagatctcgt ccatggtgct gaccaagatg aaggagatcg ccgaggcgta cctgggctac
     1681 ccggtgacca acgcggtgat caccgtgccg gcctacttca acgactcgca gcgccaggcc
     1741 accaaggatg cgggtgtgat cgcggggctc aacgtgctgc ggatcatcaa cgagcccacg
     1801 gccgccgcca tcgcctacgg cctggacaga acgggcaagg gggagcgcaa cgtgctcatc
     1861 tttgacctgg gcgggggcac cttcgacgtg tccatcctga cgatcgacga cggcatcttc
     1921 gaggtgaagg ccacggccgg ggacacccac ctgggtgggg aggactttga caacaggctg
     1981 gtgaaccact tcgtggagga gttcaagaga aaacacaaga aggacatcag ccagaacaag
     2041 cgagccgtga ggcggctgcg caccgcctgc gagagggcca agaggaccct gtcgtccagc
     2101 acccaggcca gcctggagat cgactccctg tttgagggca tcgacttcta cacgtccatc
     2161 accagggcga ggttcgagga gctgtgctcc gacctgttcc gaagcaccct ggagcccgtg
     2221 gagaaggctc tgcgcgacgc caagctggac aaggcccaga ttcacgacct ggtcctggtc
     2281 gggggctcca cccgcatccc caaggtgcag aagctgctgc aggacttctt caacgggcgc
     2341 gacctgaaca agagcatcaa ccccgacgag gctgtggcct acggggcggc ggtgcaggcg
     2401 gccatcctga tgggggacaa gtccgagaac gtgcaggacc tgctgctgct ggacgtggct
     2461 cccctgtcgc tggggctgga gacggccgga ggcgtgatga ctgccctgat caagcgcaac
     2521 tccaccatcc ccaccaagca gacgcagatc ttcaccacct actccgacaa ccaacccggg
     2581 gtgctgatcc aggtgtacga gggcgagagg gccatgacga aagacaacaa tctgttgggg
     2641 cgcttcgagc tgagcggcat ccctccggcc cccaggggcg tgccccagat cgaggtgacc
     2701 ttcgacatcg atgccaacgg catcctgaac gtcacggcca cggacaagag caccggcaag
     2761 gccaacaaga tcaccatcac caacgacaag ggccgcctga gcaaggagga gatcgagcgc
     2821 atggtgcagg aggcggagaa gtacaaagcg gaggacgagg tgcagcgcga gagggtgtca
     2881 gccaagaacg ccctggagtc ctacgccttc aacatgaaga gcgccgtgga ggatgagggg
     2941 ctcaagggca agatcagcga ggccgacaag aagaaggtgc tggacaagtg tcaagaggtc
     3001 atctcgtggc tggacgccaa caccttggcc gagaaggacg agtttgagca caagaggaag
     3061 gagctggagc aggtgtgtaa ccccatcatc agcggactgt accagggtgc cggtggtccc
     3121 gggcctgggg gcttcggggc tcagggtccc aagggagggt ctgggtcagg ccccaccatt
     3181 gaggaggtag attaggggcc tttccaagat tgctgttttt gttttggagc ttcaagactt
     3241 tgcatttcct agtatttctg tttgtcagtt ctcaatttcc tgtgtttgca atgttgaaat
     3301 tttttggtga agtcggatcc accggatcta gataactgat cataatcagc cataccacat
     3361 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3421 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3481 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3541 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     3601 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3661 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3721 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3781 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3841 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3901 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3961 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4021 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4141 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4441 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4501 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4561 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4621 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4681 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4741 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4801 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4861 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4921 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4981 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5041 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5101 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5161 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5221 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5281 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5341 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5401 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5461 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5521 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5581 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5641 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5701 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5761 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5821 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     6661 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
