LOCUS       sequence-227490-        7374 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 7374)
  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 7374)
  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: EGFP-Tubulin wt
FEATURES             Location/Qualifiers
     source          1..7374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    126..588
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(182..199)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     409..428
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             624..1427
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(687..706)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1297..1316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1459..1476)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1465..1689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1736..1755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1815..1831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1815..1831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1828..1850)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1839..1855
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1863..1869,1870..1887,1888..1893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1908..1929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2046..2063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2217..2805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2297..2316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2976..3767,3768..3836))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3599..3618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3837..3941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4016..4035)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4207..4586
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4587..4790
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4741..4761
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4835..4854
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4835..4853
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4915..4924
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4921..4923,4924..4926,4927..5637)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4921..4923,4924..4926,4927..5637)
                     /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(4966..4987)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5227..5246)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5574..5595
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          7086..7315
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     7298..7317
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 cctctccccc ccccccctct cccycncccc ccccctaacg ttactggccg aagccgcttg
       61 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
      121 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
      181 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
      241 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
      301 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
      361 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
      421 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
      481 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
      541 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac
      601 ccgggataat tcctgcagcc aatatgggat cggccattga acaagatgga ttgcacgcag
      661 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
      721 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
      781 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
      841 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
      901 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
      961 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     1021 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     1081 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     1141 tgttcgccag gctcaaggcg cgcatgcccg acggcgatga tctcgtcgtg acccatggcg
     1201 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     1261 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     1321 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     1381 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaggg gatcaattct
     1441 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1501 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1561 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1621 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     1681 gctctatggc ttctgaggcg gaaagaacca gctggggctc gagtgcattc tagttgtggt
     1741 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     1801 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1861 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1921 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1981 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2041 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2101 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2161 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2401 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2641 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3841 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3901 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3961 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     4021 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     4081 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     4141 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     4201 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     4261 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4321 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4381 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4441 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4501 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4561 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4621 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4681 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4741 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
     4801 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
     4861 aagcttggta ccgagctcgg atcgatatct gcggcctagc tagcgctacc ggtcgccacc
     4921 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4981 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     5041 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     5101 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     5161 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     5221 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     5281 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     5341 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     5401 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     5461 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5521 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5581 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
     5641 ggactcagat ctcgagtgcg tgagtgcatc tccatccacg ttggccaggc tggtgtccag
     5701 attggcaatg cctgctggga gctctactgc ctggaacacg gcatccagcc cgatggccag
     5761 atgccaagtg acaagaccat tgggggagga gatgactcct tcaacacctt cttcagtgag
     5821 acgggcgctg gcaagcacgt gccccgggct gtgtttgtag acttggaacc cacagtcatt
     5881 gatgaagttc gcactggcac ctaccgccag ctcttccacc ctgagcagct catcacaggc
     5941 aaggaagatg ctgccaataa ctatgcccga gggcactaca ccattggcaa ggagatcatt
     6001 gaccttgtgt tggaccgaat tcgcaagctg gctgaccagt gcaccggtct tcagggcttc
     6061 ttggttttcc acagctttgg tgggggaact ggttctgggt tcacctccct gctcatggaa
     6121 cgtctctcag ttgattatgg caagaagtcc aagctggagt tctccattta cccagcaccc
     6181 caggtttcca cagctgtagt tgagccctac aactccatcc tcaccaccca caccaccctg
     6241 gagcactctg attgtgcctt catggtagac aatgaggcca tctatgacat ctgtcgtaga
     6301 aacctcgata tcgagcgccc aacctacact aaccttaacc gccttattag ccagattgtg
     6361 tcctccatca ctgcttccct gagatttgat ggagccctga atgttgacct gacagaattc
     6421 cagaccaacc tggtgcccta cccccgcatc cacttccctc tggccacata tgcccctgtc
     6481 atctctgctg agaaagccta ccatgaacag ctttctgtag cagagatcac caatgcttgc
     6541 tttgagccag ccaaccagat ggtgaaatgt gaccctcgcc atggtaaata catggcttgc
     6601 tgcctgttgt accgtggtga cgtggttccc aaagatgtca atgctgccat tgccaccatc
     6661 aaaaccaagc gcagcatcca gtttgtggat tggtgcccca ctggcttcaa ggttggcatc
     6721 aactaccagc ctcccactgt ggtgcctggt ggagacctgg ccaaggtaca gagagctgtg
     6781 tgcatgctga gcaacaccac agccattgct gaggcctggg ctcgcctgga ccacaagttt
     6841 gacctgatgt atgccaagcg tgcctttgtt cactggtacg tgggtgaggg gatggaggaa
     6901 ggcgagtttt cagaggcccg tgaagatatg gctgcccttg agaaggatta tgaggaggtt
     6961 ggtgtggatt ctgttgaagg agagggtgag gaagaaggag aggaatacta aggatccgcg
     7021 gccgcataga taactgatcc agtgtgctgg aattaattcg ctgtctgcga gggccagctg
     7081 ttggggtgag tactccctct caaaagcggg catgacttct gcgctaagat tgtcagtttc
     7141 caaaaacgag gaggatttga tattcacctg gcccgcggtg atgcctttga gggtggccgc
     7201 gtccatctgg tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga
     7261 gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc
     7321 cactcccagg tccaactgca ggtcgagcat gcatctaggg cggccaattc cgcc
//
