LOCUS       sequence-343212-        8686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8686)
  TITLE     Sun Lab Overexpression Plasmids
REFERENCE   2  (bases 1 to 8686)
  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"
COMMENT     Sequence Label: pMIGR1-HA-mTBK1
FEATURES             Location/Qualifiers
     source          1..8686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     135..154
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     394..415
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    472..1024
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    474..1041
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(634..653)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     863..880
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(3265..3291)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3315..3731)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3331..3347)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3363..3385)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3798..4139
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4203..4719
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4919..4938)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5066..5088)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5282..5301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5401..5423
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5461..5479)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5547..5651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5652..5720,5721..6512)
                     /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     complement(5870..5889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6683..7271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7172..7191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7425..7442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7559..7580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7595..7600,7601..7618,7619..7625)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7633..7649
                     /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)."
     LTR             7811..8325
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(8427..8686,1..454,455..457,458..460))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8472..8493)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
       61 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
      121 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
      181 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
      241 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
      301 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
      361 cctcgccaga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
      421 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggttgtggcc atattatcat
      481 cgtgtttttc aaaggaaaac cacgtccccg tggttcgggg ggcctagacg tttttttaac
      541 ctcgactaaa cacatgtaaa gcatgtgcac cgaggcccca gatcagatcc catacaatgg
      601 ggtaccttct gggcatcctt cagccccttg ttgaatacgc ttgaggagag ccatttgact
      661 ctttccacaa ctatccaact cacaacgtgg cactggggtt gtgccgcctt tgcaggtgta
      721 tcttatacac gtggcttttg gccgcagagg cacctgtcgc caggtggggg gttccgctgc
      781 ctgcaaaggg tcgctacaga cgttgtttgt cttcaagaag cttccagagg aactgcttcc
      841 ttcacgacat tcaacagacc ttgcattcct ttggcgagag gggaaagacc cctaggaatg
      901 ctcgtcaaga agacagggcc aggtttccgg gccctcacat tgccaaaaga cggcaatatg
      961 gtggaaaata acatatagac aaacgcacac cggccttatt ccaagcggct tcggccagta
     1021 acgttagggg gggggggcgg aattgttaac agatctctaa agacagtcca cattgcgaag
     1081 gccaccatcc attgttaaag acccaaacct ttctaaaata tgattgttct cggccagctc
     1141 cttaaccacg ccttccatct cctcctttaa cttcttcata ccaagagtca tctccactaa
     1201 ggtgttagag ctggggtaga tcggggccat ggcgtgcttg acgccgccgg aggctgcgag
     1261 cattttctga ggcagagttt cttgtaactc gttagtatag tcttgatact tggacacttc
     1321 ctcttcgata tcgaaacact gattagttag cgataacagc tgcttcctaa gatgaagcat
     1381 ctttctcatc cactcttccg acttatcttt aaacgcttca tactttctaa cacattcttc
     1441 tgagaagtgg ctcattgctt ttgtggcatg gtaatacaat ttttgcttat caaatttgtg
     1501 gatctgttct tcattataag ctagtctgcg ttctgctttg tcttttttga actgatagta
     1561 aatctctgtg atgcagttca acaggacctg cagtttttct acattcctgt ctcttggatg
     1621 cgtgccttct tgatgtgccc aggtgtcggc cagcaagccc cctggagaca gcctgctgct
     1681 gatgtcctga agactgcttt ctattgttcc ctgagaactg gaaagtctca gcagcttggt
     1741 gtgtatgtct gaaatctcac ccagctctgc ggcttccagg ttgaccttca tcaacttctc
     1801 atacactttc acagtcttct caatgttcct gatgcagaaa tccagtgtga tcactacctc
     1861 cgtcttcttg tggacggtct cgttgtaatc atccttaacc agttcaacca gccaccgtac
     1921 cccctttcgc attaattctt gatagagcag cagggtactg gcagttctgc aggcgtagca
     1981 cacaaccccc gtcactgcct tggccatgct ggcgtcccca tccagatcat agcgtggatg
     2041 tattttaggg agggaaattt tttcatatct cagtcctacg gtattgagtt gttcccggct
     2101 cgtgacaaag ataggatttt cctctgtggt tttaggaaaa tgctgggcta gtcgtccgag
     2161 ttctaggact aagcgtcgtc cttcgtagat aagttcttga tttgaggaaa caatcttggt
     2221 ttgtttatag accagttcat ggaacacagc agcagtgtta tagctgtgaa tgtaaatctt
     2281 atgcgccgtc atgtgttgta gcgagaagac atggatcacc attcggtgaa gcacatcact
     2341 ggtctctgca aagaactggt caaaacccca gcacttctcc tgatcagctt caagtatgtt
     2401 tgcaagaact ggggtaagca gtgcctgaag accctgagaa agactacagg agagaggcat
     2461 gtctccactc cagtcaattg gtccgttttc tgctttctgt actccagata ttgcaccaga
     2521 cggcttccca gtgattattt tatacattac ttctttgttc ctccgaggcc cctcgaaggg
     2581 tctaaacggc agcgaccccg tggctgcatg gtagaatgtc actccaacac tccacagatc
     2641 aacggtagcc ccgtacttct tctgatggtc ctttcttagc actgcccttt catacatgtc
     2701 cggatgcagg tactcttctg tgccgtacag agacacaaac tgctcatcgt cctccagctc
     2761 tcgagcggcg ccgaaatccg tgagtttgta cacagactgg ccgtcctccc ctatgacgcg
     2821 catgatgttg cctggcttga tatctcggtg cactatgccg ttctctcgga gatgattcat
     2881 cccgcccacc acatctcgta agacaatgag aaattctgat tctggaagtc catacgcatt
     2941 ggacggctcc tctagaacag tgtataaact cccacaggga caaaactcca taataagcac
     3001 tttatgtctt gttgttgtct cctcttcaat agcaaataac ttgacaatgt ttttgtgatt
     3061 gagttttttt aacacttcaa attctctcat ttgaacatcc actgggcgaa ggaagcttat
     3121 gttattaaat actttgacag catagagatc accagttttc ttatgccttc ctcggaagac
     3181 atttgcagtg gccccctggc ctaggatatc agacaggagc cacagatggt tggaggtgct
     3241 ctgcatggtg aattcgcgct gagcagcgta atctggaacg tcatatggat agcccatctc
     3301 gaggatctaa ttccggcgcc tagagaagga gtgagggctg gataaaggga ggatcgaggc
     3361 ggggtcgaac gaggaggttc aagggggaga gacggggcgg atggaggaag aggaggcgga
     3421 ggcttagggt gtacaaaggg cttgacccag ggaggggggt caaaagccaa ggcttcccag
     3481 gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc caggtgaaaa gaccttgatc
     3541 ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc gcggccatcc gacgttaaag
     3601 gttggccatt ctgcagagca gaaggtaacc caacgtctct tcttgacatc taccgactgg
     3661 ttgtgagcga tccgctcgac atctttccag tgacctaagg tcaaacttaa gggagtggta
     3721 acagtctggc cctaattttc agacaaatac agaaacacag tcagacagag acaacacaga
     3781 acgatgctgc agcgctgcag cagacaagac gcgcggcttc ggttccaaac cgaaagcaaa
     3841 aattcagacg gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac cacatatcct
     3901 gacggggtcg gattccacat cgactccctt cctcaggtcg ggccacaaaa acggccccca
     3961 aagtccctgg gacgtctccc agggttgcgg ccgggtgttc agaactcgtc agttccacca
     4021 cgggtccgcc agatacagag ctagttagct aactagtaca gacgcaggcg caaacattag
     4081 atgccggcac aaacacgcac aaagacagag acagacacga aacgaccgct ggccagctta
     4141 cctcccggcg ggggggtcgg tggtccctgg gcaggggtct ccaaatctcg gtggaacctc
     4201 caaatgaaag acccccgagg tgggcagtca atcaatctga ggagaccctc ccaaggatca
     4261 gcgagtccac gattcggatg caaacagcaa gaggctttat tgggaatacg ggtacccggg
     4321 cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag
     4381 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata
     4441 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa
     4501 actgctgagg gcgggaccgc atctggggac catctgttct tggccctgag ccggggcagg
     4561 aactgcttac cacagatatc ctgtttggcc catattctgc tgtctctctg ttcctaacct
     4621 tgatctgaac ttctctattc tcagttatgt attttccatg ccttgcaaaa tggcgttact
     4681 taagctagct tgccaaacct acaggtgggg tctttcattc cccccttttt ctggagacta
     4741 aataaaatct tttattttat ctatggctcg tactctatag gcttcagctg gtgatattgt
     4801 tgagtcaaaa ctagagcctg gaccactgat atcctgtctt taacaaattg gactaatcgc
     4861 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     4921 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     4981 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     5041 ttgcatgcac cattccttgc gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5101 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5161 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     5221 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     5281 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5341 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5401 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5461 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5521 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5581 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5641 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5701 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5761 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5821 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5881 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5941 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6001 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6061 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6121 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6181 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6241 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6301 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6361 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6421 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6481 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6541 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6601 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6661 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6721 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6781 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6841 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6901 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6961 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7021 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7081 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7141 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7201 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7261 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7321 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7381 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7441 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7501 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7561 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7621 tgttgtgtgg aattgtgagc ggataacaat ttcacaattc tccccagctc tttctgcgct
     7681 gtccataatg aactatttca ggagtattgg agtatgtgga ttcaaaacag tggctaattg
     7741 agtcaggatt acttaatatt cgactcctac cctcagaatg ttgttctcca acttcaagaa
     7801 actgttaccc atgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc
     7861 aaggaacagc gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt
     7921 acccgggcga cgcagtctat cggaggactg gcgcgccgag tgaggggttg tgggctcttt
     7981 tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt
     8041 tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc
     8101 tctagaaact gctgagggcg ggaccgcatc tggggaccat ctgttcttgg ccctgagccg
     8161 gggcaggaac tgcttaccac agatatcctg tttggcccat attctgctgt ctctctgttc
     8221 ctaaccttga tctgaacttc tctattctca gttatgtatt ttccatgcct tgcaaaatgg
     8281 cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc cctttttctg
     8341 gagactaaat aaaatctttt attttatcga taagcttggc tgcaggtcga cggtatcgat
     8401 aagcttgata attcttaatt aattcattac ttgtacagct cgtccatgcc gagagtgatc
     8461 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     8521 ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     8581 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     8641 cggatcttga agttcgcctt gatgccgttc ttctgcttgt cggcca
//
