LOCUS       sequence-239403-        7816 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 7816)
  AUTHORS   Gao YS, Hubbert CC, Yao TP
  TITLE     The microtubule-associated histone deacetylase 6 (HDAC6) regulates 
            epidermal growth factor receptor (EGFR) endocytic trafficking and 
            degradation.
  JOURNAL   J Biol Chem. 2010 Apr 9. 285(15):11219-26.
  PUBMED    20133936
REFERENCE   2  (bases 1 to 7816)
  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. 2010 Apr 9. 285(15):11219-26."
COMMENT     Sequence Label: pEGFP.N1-HDAC6.delta Buz
FEATURES             Location/Qualifiers
     source          1..7816
                     /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"
     regulatory      3756..3765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3762..3764,3765..3767,3768..4481)
                     /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(3762..3764,3765..3767,3768..4478)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3807..3828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4068..4087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4415..4436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4604..4725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4641..4660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4695..4714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4732..5187)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4869..4890)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5081..5100
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5214..5318
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5320..5677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5320..5511
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5343..5363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5528..5663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5712..6506
                     /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             5712..6506
                     /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(5766..5785)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6376..6395
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6694..6713)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6738..6785
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7114..7702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7603..7622
                     /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 atctatgacc tcaaccggcc aggattccac cacaaccagg cagcgaagaa
      661 gtaggcagaa cccccagtcg ccccctcagg actccagtgt cacttcgaag cgaaatatta
      721 aaaagggagc cgttccccgc tctatcccca atctagcgga ggtaaagaag aaaggcaaaa
      781 tgaagaagct cggccaagca atggaagaag acctaatcgt gggactgcaa gggatggatc
      841 tgaaccttga ggctgaagca ctggctggca ctggcttggt gttggatgag cagttaaatg
      901 aattccattg cctctgggat gacagcttcc cggaaggccc tgagcggctc catgccatca
      961 aggagcaact gatccaggag ggcctcctag atcgctgcgt gtcctttcag gcccggtttg
     1021 ctgaaaagga agagctgatg ttggttcaca gcctagaata tattgatctg atggaaacaa
     1081 cccagtacat gaatgaggga gaactccgtg tcctagcaga cacctacgac tcagtttatc
     1141 tgcatccgaa ctcatactcc tgtgcctgcc tggcctcagg ctctgtcctc aggctggtgg
     1201 atgcggtcct gggggctgag atccggaatg gcatggccat cattaggcct cctggacatc
     1261 acgcccagca cagtcttatg gatggctatt gcatgttcaa ccacgtggct gtggcagccc
     1321 gctatgctca acagaaacac cgcatccgga gggtccttat cgtagattgg gatgtgcacc
     1381 acggtcaagg aacacagttc accttcgacc aggaccccag tgtcctctat ttctccatcc
     1441 accgctacga gcagggtagg ttctggcccc acctgaaggc ctctaactgg tccaccacag
     1501 gtttcggcca aggccaagga tataccatca atgtgccttg gaaccaggtg gggatgcggg
     1561 atgctgacta cattgctgct ttcctgcacg tcctgctgcc agtcgccctc gagttccagc
     1621 ctcagctggt cctggtggct gctggatttg atgccctgca aggggacccc aagggtgaga
     1681 tggccgccac tccggcaggg ttcgcccagc taacccacct gctcatgggt ctggcaggag
     1741 gcaagctgat cctgtctctg gagggtggct acaacctccg cgccctggct gaaggcgtca
     1801 gtgcttcgct ccacaccctt ctgggagacc cttgccccat gctggagtca cctggtgccc
     1861 cctgccggag tgcccaggct tcagtttcct gtgctctgga agcccttgag cccttctggg
     1921 aggttcttgt gagatcaact gagaccgtgg agagggacaa catggaggag gacaatgtag
     1981 aggagagcga ggaggaagga ccctgggagc cccctgtgct cccaatcctg acatggccag
     2041 tgctacagtc tcgcacaggg ctggtctatg accaaaatat gatgaatcac tgcaacttgt
     2101 gggacagcca ccaccctgag gtaccccagc gcatcttgcg gatcatgtgc cgtctggagg
     2161 agctgggcct tgccgggcgc tgcctcaccc tgacaccgcg ccctgccaca gaggctgagc
     2221 tgctcacctg tcacagtgct gagtacgtgg gtcatctccg ggccacagag aaaatgaaaa
     2281 cccgggagct gcaccgtgag agttccaact ttgactccat ctatatctgc cccagtacct
     2341 tcgcctgtgc acagcttgcc actggcgctg cctgccgcct ggtggaggct gtgctctcag
     2401 gagaggttct gaatggtgct gctgtggtgc gtcccccagg acaccacgca gagcaggatg
     2461 cagcttgcgg tttttgcttt ttcaactctg tggctgtggc tgctcgccat gcccagacta
     2521 tcagtgggca tgccctacgg atcctgattg tggattggga tgtccaccac ggtaatggaa
     2581 ctcagcacat gtttgaggat gaccccagtg tgctatatgt gtccctgcac cgctatgatc
     2641 atggcacctt cttccccatg ggggatgagg gtgccagcag ccagatcggc cgggctgcgg
     2701 gcacaggctt caccgtcaac gtggcatgga acgggccccg catgggtgat gctgactacc
     2761 tagctgcctg gcatcgcctg gtgcttccca ttgcctacga gtttaaccca gaactggtgc
     2821 tggtctcagc tggctttgat gctgcacggg gggatccgct ggggggctgc caggtgtcac
     2881 ctgagggtta tgcccacctc acccacctgc tgatgggcct tgccagtggc cgcattatcc
     2941 ttatcctaga gggtggctat aacctgacat ccatctcaga gtccatggct gcctgcactc
     3001 gctccctcct tggagaccca ccacccctgc tgaccctgcc acggccccca ctatcagggg
     3061 ccctggcctc aatcactgag accatccaag tccatcgcag atactggcgc agcttacggg
     3121 tcatgaaggt agaagacaga gaaggaccct ccagttctaa gttggtcacc aagaaggcac
     3181 cccaaccagc caaacctagg ttagctgagc ggatgaccac acgagaaaag aaggttctgg
     3241 aagcaggcat ggggaaagtc acctcggcat catttgggga agagtccact ccaggccaga
     3301 ctaactcaga gacagctgtg gtggccctca ctcaggacca gccctcagag gcagccacag
     3361 ggggagccac tctggcccag accatttctg aggcagccat tgggggagcc atgctgggcc
     3421 agaccacctc agaggaggct gtcgggggag ccactccgga ccagaccacc tcagaggaga
     3481 ctgtgggagg agccattctg gaccagacca cctcagagga tgctgttggg ggagccacgc
     3541 tgggccagac tacctcagag gaggctgtag gaggagctac actggcccag accatctcgg
     3601 aggcagccat ggagggagcc acactggacc agactacgtc agaggaggct ccagggggca
     3661 ccgagctgat ccaaactcct ctagcctcga gcacagacca ccagaccccc ccaacctcac
     3721 ctgtgcaggg aactacaccc cagatgggat ccatcgccac catggtgagc aagggcgagg
     3781 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3841 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3901 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3961 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4021 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4081 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4141 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4201 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4261 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4321 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4381 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4441 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat
     4501 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4561 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4621 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4681 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     4741 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4801 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4861 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4921 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4981 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5041 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5101 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5161 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     5341 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5401 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5461 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5521 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5581 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5641 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     5701 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5761 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5821 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5881 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5941 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6001 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     6061 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     6121 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     6181 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     6241 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     6301 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     6361 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6421 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6481 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6541 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6601 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6661 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6721 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     6781 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6841 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6901 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6961 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7021 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7261 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7801 tattaccgcc atgcat
//
