LOCUS       sequence-159448-        8371 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 8371)
  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 8371)
  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.DC (catalytic deficient)
FEATURES             Location/Qualifiers
     source          1..8371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4266..4275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4272..4274,4275..4277,4278..4991)
                     /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(4272..4274,4275..4277,4278..4988)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4317..4338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4578..4597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4925..4946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5114..5235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5151..5170)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5205..5224
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5242..5697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5379..5400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5591..5610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5724..5828
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5830..6187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5830..6021
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5853..5873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6038..6173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6222..7016
                     /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             6222..7016
                     /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(6276..6295)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6886..6905
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7204..7223)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7248..7295
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7624..8212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8113..8132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatcta tgacctcaac cggccaggat tccaccacaa
      601 ccaggcagcg aagaagtagg cagaaccccc agtcgccccc tcaggactcc agtgtcactt
      661 cgaagcgaaa tattaaaaag ggagccgttc cccgctctat ccccaatcta gcggaggtaa
      721 agaagaaagg caaaatgaag aagctcggcc aagcaatgga agaagaccta atcgtgggac
      781 tgcaagggat ggatctgaac cttgaggctg aagcactggc tggcactggc ttggtgttgg
      841 atgagcagtt aaatgaattc cattgcctct gggatgacag cttcccggaa ggccctgagc
      901 ggctccatgc catcaaggag caactgatcc aggagggcct cctagatcgc tgcgtgtcct
      961 ttcaggcccg gtttgctgaa aaggaagagc tgatgttggt tcacagccta gaatatattg
     1021 atctgatgga aacaacccag tacatgaatg agggagaact ccgtgtccta gcagacacct
     1081 acgactcagt ttatctgcat ccgaactcat actcctgtgc ctgcctggcc tcaggctctg
     1141 tcctcaggct ggtggatgcg gtcctggggg ctgagatccg gaatggcatg gccatcatta
     1201 ggcctcctgg acatgccgcc cagcacagtc ttatggatgg ctattgcatg ttcaaccacg
     1261 tggctgtggc agcccgctat gctcaacaga aacaccgcat ccggagggtc cttatcgtag
     1321 attgggatgt gcaccacggt caaggaacac agttcacctt cgaccaggac cccagtgtcc
     1381 tctatttctc catccaccgc tacgagcagg gtaggttctg gccccacctg aaggcctcta
     1441 actggtccac cacaggtttc ggccaaggcc aaggatatac catcaatgtg ccttggaacc
     1501 aggtggggat gcgggatgct gactacattg ctgctttcct gcacgtcctg ctgccagtcg
     1561 ccctcgagtt ccagcctcag ctggtcctgg tggctgctgg atttgatgcc ctgcaagggg
     1621 accccaaggg tgagatggcc gccactccgg cagggttcgc ccagctaacc cacctgctca
     1681 tgggtctggc aggaggcaag ctgatcctgt ctctggaggg tggctacaac ctccgcgccc
     1741 tggctgaagg cgtcagtgct tcgctccaca cccttctggg agacccttgc cccatgctgg
     1801 agtcacctgg tgccccctgc cggagtgccc aggcttcagt ttcctgtgct ctggaagccc
     1861 ttgagccctt ctgggaggtt cttgtgagat caactgagac cgtggagagg gacaacatgg
     1921 aggaggacaa tgtagaggag agcgaggagg aaggaccctg ggagccccct gtgctcccaa
     1981 tcctgacatg gccagtgcta cagtctcgca cagggctggt ctatgaccaa aatatgatga
     2041 atcactgcaa cttgtgggac agccaccacc ctgaggtacc ccagcgcatc ttgcggatca
     2101 tgtgccgtct ggaggagctg ggccttgccg ggcgctgcct caccctgaca ccgcgccctg
     2161 ccacagaggc tgagctgctc acctgtcaca gtgctgagta cgtgggtcat ctccgggcca
     2221 cagagaaaat gaaaacccgg gagctgcacc gtgagagttc caactttgac tccatctata
     2281 tctgccccag taccttcgcc tgtgcacagc ttgccactgg cgctgcctgc cgcctggtgg
     2341 aggctgtgct ctcaggagag gttctgaatg gtgctgctgt ggtgcgaccc ccaggacacg
     2401 ccgcagagca ggatgcagct tgcggttttt gctttttcaa ctctgtggct gtggctgctc
     2461 gccatgccca gactatcagt gggcatgccc tacggatcct gattgtggat tgggatgtcc
     2521 accacggtaa tggaactcag cacatgtttg aggatgaccc cagtgtgcta tatgtgtccc
     2581 tgcaccgcta tgatcatggc accttcttcc ccatggggga tgagggtgcc agcagccaga
     2641 tcggccgggc tgcgggcaca ggcttcaccg tcaacgtggc atggaacggg ccccgcatgg
     2701 gtgatgctga ctacctagct gcctggcatc gcctggtgct tcccattgcc tacgagttta
     2761 acccagaact ggtgctggtc tcagctggct ttgatgctgc acggggggat ccgctggggg
     2821 gctgccaggt gtcacctgag ggttatgccc acctcaccca cctgctgatg ggccttgcca
     2881 gtggccgcat tatccttatc ctagagggtg gctataacct gacatccatc tcagagtcca
     2941 tggctgcctg cactcgctcc ctccttggag acccaccacc cctgctgacc ctgccacggc
     3001 ccccactatc aggggccctg gcctcaatca ctgagaccat ccaagtccat cgcagatact
     3061 ggcgcagctt acgggtcatg aaggtagaag acagagaagg accctccagt tctaagttgg
     3121 tcaccaagaa ggcaccccaa ccagccaaac ctaggttagc tgagcggatg accacacgag
     3181 aaaagaaggt tctggaagca ggcatgggga aagtcacctc ggcatcattt ggggaagagt
     3241 ccactccagg ccagactaac tcagagacag ctgtggtggc cctcactcag gaccagccct
     3301 cagaggcagc cacaggggga gccactctgg cccagaccat ttctgaggca gccattgggg
     3361 gagccatgct gggccagacc acctcagagg aggctgtcgg gggagccact ccggaccaga
     3421 ccacctcaga ggagactgtg ggaggagcca ttctggacca gaccacctca gaggatgctg
     3481 ttgggggagc cacgctgggc cagactacct cagaggaggc tgtaggagga gctacactgg
     3541 cccagaccat ctcggaggca gccatggagg gagccacact ggaccagact acgtcagagg
     3601 aggctccagg gggcaccgag ctgatccaaa ctcctctagc ctcgagcaca gaccaccaga
     3661 cccccccaac ctcacctgtg cagggaacta caccccagat atctcccagt acactgattg
     3721 ggagtctcag gaccttggag ctaggcagcg aatctcaggg ggcctcagaa tctcaggccc
     3781 caggagagga gaacctacta ggagaggcag ctggaggtca ggacatggct gattcgatgc
     3841 tgatgcaggg atctaggggc ctcactgatc aggccatatt ttatgctgtg acaccactgc
     3901 cctggtgtcc ccatttggtg gcagtatgcc ccatacctgc agcaggccta gacgtgaccc
     3961 aaccttgtgg ggactgtgga acaatccaag agaattgggt gtgtctctct tgctatcagg
     4021 tctactgtgg tcgttacatc aatggccaca tgctccaaca ccatggaaat tctggacacc
     4081 cgctggtcct cagctacatc gacctgtcag cctggtgtta ctactgtcag gcctatgtcc
     4141 accaccaggc tctcctagat gtgaagaaca tcgcccacca gaacaagttt ggggaggata
     4201 tgccccaccc acacaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     4261 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     4321 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     4381 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     4441 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     4501 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     4561 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     4621 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4681 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     4741 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     4801 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4861 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     4921 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     4981 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     5041 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5101 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5161 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5221 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5281 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5341 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5401 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5461 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5521 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5581 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5641 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     5701 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5761 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5821 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     5881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5941 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6001 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6061 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6121 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6181 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     6241 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6301 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6361 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     6421 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6481 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6541 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6601 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6661 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6721 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     6781 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6841 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6901 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6961 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     7021 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     7081 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7141 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     7201 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     7261 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     7321 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     7381 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     7441 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     7501 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7561 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7621 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7681 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7741 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7801 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7861 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7921 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7981 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8041 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8101 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8161 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8221 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8281 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     8341 aatcaattac ggggtcatta gttcatagcc c
//
