LOCUS       sequence-343670-        8232 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 8232)
  TITLE     Sun Lab Overexpression Plasmids
REFERENCE   2  (bases 1 to 8232)
  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-mTak1
FEATURES             Location/Qualifiers
     source          1..8232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    165..506
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             573..989
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     919..941
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     957..973
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1013..1039
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1873..1884)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2809..3376
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2826..3378
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2970..2987)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3197..3216
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3390..3392,3393..3395,3396..4109)
                     /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(3435..3456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3696..3715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4043..4064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4211..4725
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4887..4903
                     /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(4911..4917,4918..4935,4936..4941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4956..4977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5094..5111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5265..5853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5345..5364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6024..6815,6816..6884))
                     /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     6647..6666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6885..6989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7057..7075
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7113..7135)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7235..7254
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7448..7470
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7598..7617
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7817..8232,1..101)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
       61 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
      121 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
      181 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
      241 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
      301 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
      361 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
      421 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
      481 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
      541 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
      601 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
      661 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
      721 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
      781 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
      841 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
      901 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
      961 atccagccct cactccttct ctaggcgccg gaattagatc ctcgagatgg gctatccata
     1021 tgacgttcca gattacgctg ctcagcgcga attcatgtcc acagcctccg ccgcctcgtc
     1081 ctcctcctcg tcttctgcca gtgagatgat cgaagcgccg tcgcaggtcc tgaacttcga
     1141 agagatcgac tacaaggaga tcgaggtgga agaggttgtc ggaagaggag cttttggagt
     1201 agtttgcaaa gctaagtgga gagcaaaaga tgtcgctatt aaacagatag aaagtgagtc
     1261 tgagaggaag gctttcattg tggagctccg gcagttgtcg cgtgtgaacc atcctaacat
     1321 tgtcaagttg tacggagcct gcctgaatcc agtatgtctt gtgatggaat atgcagaggg
     1381 gggctcattg tataatgtgc tgcatggtgc tgaaccattg ccttactaca ctgctgctca
     1441 tgccatgagc tggtgtttac agtgttccca aggagtggct tacctgcaca gcatgcagcc
     1501 caaagcgctg attcacaggg acctcaagcc tccaaacttg ctgctggttg caggagggac
     1561 agttctaaaa atctgcgatt ttggtacagc ttgtgacatc caaacacaca tgaccaataa
     1621 taaagggagt gctgcttgga tggcgcctga agtgtttgaa ggtagcaatt acagtgaaaa
     1681 gtgtgatgtc ttcagctggg gtattatcct ctgggaagtg ataacacgcc ggaaaccctt
     1741 cgatgagatc ggtggcccag ctttcagaat catgtgggct gttcataatg gcactcgacc
     1801 accactgatc aaaaatttac ctaagcccat tgagagcttg atgacacgct gttggtctaa
     1861 ggacccatct cagcgccctt caatggagga aattgtgaaa ataatgactc acttgatgcg
     1921 gtacttccca ggagcggatg agccattaca gtatccttgt cagtactctg atgaagggca
     1981 gagcaactca gccaccagca caggctcgtt catggacatt gcttctacaa ataccagtaa
     2041 taaaagtgac acaaatatgg aacaggttcc tgccacaaac gacactatta aacgcttgga
     2101 gtcaaaactg ttgaaaaacc aggcaaagca acagagtgaa tctggacgcc tgagcttggg
     2161 agcctctcgt gggagcagtg tggagagctt gccccccact tccgagggca agaggatgag
     2221 tgctgacatg tctgaaatag aagccaggat cgtggcgact gcaggtaacg ggcaaccaag
     2281 gcgtagatcc atccaagact tgactgttac tgggacagaa cctggtcagg tgagcagccg
     2341 gtcatccagc cctagtgtca gaatgatcac tacctcagga ccaacctcag agaagccagc
     2401 tcgcagtcac ccatggaccc ctgatgattc cacagacacc aatggctcag ataactccat
     2461 cccaatggcg tatcttacac tggatcacca gctacagcct ctagcgccgt gcccaaactc
     2521 caaagaatcc atggcagtgt tcgaacagca ctgtaaaatg gcacaggagt atatgaaagt
     2581 tcaaaccgaa atcgcattgt tactacagag aaagcaagaa ctagttgcag aattggacca
     2641 ggatgaaaag gaccagcaaa atacatctcg tctggtacag gaacataaaa agcttttaga
     2701 tgaaaacaaa agcctttcta cttattacca gcaatgcaaa aaacaactag aggtcatcag
     2761 aagccaacag cagaaacgac aaggcacttc atgagatctg ttaacaattc cgcccccccc
     2821 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     2881 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     2941 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3001 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3061 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3121 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3181 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3241 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     3301 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3361 aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg
     3421 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3481 ctggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3541 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3601 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3661 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3721 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3781 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3841 tctatatcat ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca
     3901 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3961 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4021 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4081 ctctcggcat ggacgagctg tacaagtaat gaattaatta agaattatca agcttatcga
     4141 taccgtcgac ctgcagccaa gcttatcgat aaaataaaag attttattta gtctccagaa
     4201 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     4261 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     4321 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4381 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     4441 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     4501 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     4561 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     4621 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     4681 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     4741 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     4801 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     4861 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     4921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7021 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7081 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     7141 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     7201 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7261 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7321 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7381 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7441 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga
     7501 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     7561 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     7621 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat
     7681 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     7741 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc
     7801 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     7861 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     7921 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     7981 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     8041 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     8101 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     8161 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     8221 gtacccgtat tc
//
