LOCUS       sequence-77205-e        6305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of myc-tagged human MID2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  AUTHORS   Short KM, Hopwood B, Yi Z, Cox TC
  TITLE     MID1 and MID2 homo- and heterodimerise to tether the 
            rapamycin-sensitive PP2A regulatory subunit, alpha 4, to 
            microtubules: implications for the clinical variability of X-linked 
            Opitz GBBB syndrome and other developmental disorders.
  JOURNAL   BMC Cell Biol. 2002;3:1. Epub 2002 Jan 4.
  PUBMED    11806752
REFERENCE   2  (bases 1 to 6305)
  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: "BMC Cell 
            Biol."; date: "2002"; volume: "3"; pages: "1. Epub 2002 Jan 4"
COMMENT     Sequence Label: pCMV-N2-6MYC-hMID2
FEATURES             Location/Qualifiers
     source          1..6305
                     /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"
     CDS             634..663
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             670..699
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             706..735
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             754..783
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             790..819
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             826..855
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3093..3214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3130..3149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3184..3203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3221..3676)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3358..3379)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3570..3589
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3703..3807
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3809..4166
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3809..4000
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3832..3852)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4017..4152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4201..4995
                     /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             4201..4995
                     /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(4255..4274)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4865..4884
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5183..5202)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5227..5274
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5603..6191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6092..6111
                     /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 atctcgagct caagcttatg ggtgaacaaa agttgatttc tgaagaagat
      661 ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc
      721 tcagaagaag acttgaacgg atcctctaga ggtgaacaaa agttgatttc tgaagaagat
      781 ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc
      841 tcagaagaag acttgaaccc aagcttcgaa ttcctgaaga tggaaacact ggagtctgaa
      901 ttgacctgtc caatctgcct agagttgttt gaagaccccc ttctgctccc ttgtgctcac
      961 agcctctgct tcagctgtgc ccatcgcatt ttggtatcaa gctgcagctc tggtgaatcc
     1021 attgaaccca ttactgcttt ccagtgtcct acctgcaggt atgttatctc gctgaaccac
     1081 cggggcctgg atggcctcaa gaggaatgtg actctgcaga acattattga tcgcttccag
     1141 aaggcttcag tcagtgggcc caattcccct agtgagagcc gccgggaaag gacttacagg
     1201 cccaccactg ccatgtctag cgagcgaatt gcttgccaat tctgtgagca ggacccgcca
     1261 agggatgcag taaaaacatg catcacctgt gaggtctcct actgtgaccg ttgcctgcgg
     1321 gccacgcacc ccaacaagaa acctttcacc agccaccgcc tggtggaacc agtgccagac
     1381 acacatcttc gagggatcac ctgcctggac catgagaatg agaaagtgaa catgtactgt
     1441 gtatctgatg accaattgat ctgtgcctta tgcaaactgg tgggtcgtca ccgagaccat
     1501 caggtcgcat ccctgaatga tcgatttgag aaactcaagc aaactctgga gatgaacctc
     1561 accaacctgg ttaagcgcaa cagcgaacta gaaaatcaaa tggccaaact aatacagatc
     1621 tgccagcagg ttgaggtgaa tactgctatg catgaggcaa aacttatgga agaatgtgac
     1681 gagttggtag agatcatcca gcagaggaag caaatgatcg ctgtcaaaat caaagagaca
     1741 aaggttatga aactgagaaa gttggcacag caggttgcta attgccgcca gtgtcttgaa
     1801 cggtcaacag tcctcatcaa ccaagctgag catatcctga aagaaaatga ccaggcacgg
     1861 tttctacagt ctgcaaaaaa tattgctgag agggtcgcta tggcaactgc atcttctcaa
     1921 gttctgattc cagacatcaa ttttaatgat gcctttgaaa actttgcttt agatttttcc
     1981 agagaaaaga aactgctaga ggggttagat tatttaacag ccccaaaccc accatctatc
     2041 cgagaagaac tctgtactgc ctcccatgac accattacag tccactggat ctcggatgat
     2101 gagttcagca tcagctccta tgagcttcag tacaccatat tcactggcca ggctaacttc
     2161 atcagcctgt ataattcagt agacagctgg atgattgttc ccaacattaa acagaaccat
     2221 tacacagtgc atggactcca gagcgggact cgctacatct tcatcgttaa agccataaac
     2281 caagccggca gccggaacag tgaacctacc cgactaaaaa caaacagcca accctttaaa
     2341 ttggatccca aaatgactca caagaagttg aagatctcca atgatggatt gcagatggag
     2401 aaggatgaaa gctctctaaa gaagagccac accccagaga ggtttagtgg cacagggtgc
     2461 tatggggcag caggaaatat attcattgac agtggctgcc actattggga ggtggtcatg
     2521 ggttcctcaa catggtatgc aattggcatt gcctacaaat cagctccaaa gaatgaatgg
     2581 attggcaaga atgcctcctc atgggtcttc tctcgctgca atagtaactt cgtggtgaga
     2641 cacaacaaca aggaaatgct ggtggatgtg cccccacacc tgaagcgtct gggtgtcctc
     2701 ctggattatg acaacaatat gctgtctttc tatgacccag ctaactctct ccatcttcat
     2761 acttttgatg tgaccttcat tcttccagtt tgtccaacat ttacaatctg gaacaaatcc
     2821 ctaatgatcc tgtctggctt gcctgcccca gattttattg attaccctga gcggcaggaa
     2881 tgcaactgca ggcctcaaga atccccttat gtttctggga tgaaaacctg tcattaagaa
     2941 ttctgcagtc gacggtaccg cgggcccggg atcggccgcg actctagatc ataatcagcc
     3001 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3061 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3121 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3181 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     3241 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3301 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3361 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3421 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3481 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3541 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3601 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3661 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3721 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3781 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3841 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3901 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3961 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4021 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4081 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4141 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4201 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4261 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4321 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4381 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4441 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4501 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4561 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4621 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4681 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4741 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4801 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4861 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4921 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4981 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5041 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5101 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5161 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5221 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5281 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5341 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5401 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6301 tgcat
//
