LOCUS       sequence-479877-        8438 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expresses Gab2 (mouse) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  AUTHORS   Kramer MH, Richardson SN, Li Y, Yin T, Helton NM, George DR, Cai M, 
            Ramakrishnan SM, Katerndahl CDS, Miller CA, Ley TJ
  TITLE     Overexpression of the signaling-coordinator GAB2 can play an 
            important role in Acute Myeloid Leukemia progression
  JOURNAL   J Clin Invest. 2025.
REFERENCE   2  (bases 1 to 8438)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Clin 
            Invest. 2025."
COMMENT     Sequence Label: Gab2-MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(116..220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     288..306
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(344..366)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     466..485
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     679..701
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     829..848
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             1048..1564
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1628..1969
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2036..2452
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2382..2404
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2420..2436
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4478..5045
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4495..5047
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4639..4656)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4866..4885
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5059..5061,5062..5064,5065..5778)
                     /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(5104..5125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5365..5384)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5712..5733
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5880..6394
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6556..6572
                     /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(6580..6586,6587..6604,6605..6610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6625..6646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6763..6780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6934..7522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7014..7033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7693..8438,1..46,47..115))
                     /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     8316..8335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
       61 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      121 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      181 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      241 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
      301 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      361 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      421 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      481 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      541 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      601 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      661 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
      721 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
      781 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
      841 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      901 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
      961 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     1021 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     1081 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     1141 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     1201 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     1261 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     1321 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     1381 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     1441 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     1501 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
     1561 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc
     1621 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc
     1681 cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     1741 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     1801 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     1861 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     1921 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     1981 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     2041 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     2101 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     2161 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     2221 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     2281 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     2341 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     2401 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag
     2461 atctcatgag cggcggcggc ggcgacgacg tggtgtgtac cggctggctg aggaaatcgc
     2521 ctcccgagaa gaagttgagg cgctatgcct ggaagaaacg ctggtttata cttcggagtg
     2581 gccgaatgag tggagatcca gatgttctgg aatactacaa gaatgagcac tccaagaaac
     2641 ccctgcggat catcaacctg aacttgtgtg agcaggtgga tgcaggcctg accttcaaca
     2701 agaaagagct gcaggatagt tttgtgtttg atatcaagac cagcgagcgc acattttacc
     2761 tggtggctga gacagaggct gacatgaata agtgggtcca gagcatctgc cagatctgcg
     2821 gcttcaatca ggctgaagag agcacagact ccctgaggaa cctttcttca gccagtcatg
     2881 gtccccgctc ttctccagct gagttcagct ccagtcagca cctgctccga gaacggaagt
     2941 cctcagcccc ttcacactct agccagccta ctttattcac gtttgagccc cctgtgtcaa
     3001 gccacatgca gcctaccttg tccaccagtg cacctcagga gtatctctac ttgcaccagt
     3061 gcataagcag aaggacagaa aatgcaagga gtgccagctt ctctcagggc acccggcaga
     3121 agagtgatac agctgtgcaa aaacttgccc agagcaatgg acactgtatc aacggcgtcg
     3181 gaggtcaagt ccatggcttc tatagccttc ccaagccaag ccgacacaat acagaattca
     3241 aagacagtac ttatgatctc ccacggagcc tggcttccca tggccacacc aagagcagcc
     3301 tcacagggtc tgagactgat aacgaggatg tgtacacctt caagatgccc agcaacaccc
     3361 tgtgtcggga acttggagac ctccttgtgg acaatatgga tgtcccaacc actcctctct
     3421 cagcctacca gatccctaga acattcacac tggacaagaa ccacaatgcc atgacagtgg
     3481 ccactcctgg agattcagcc atagctcccc caccccggcc acccaagcca agtcaggcag
     3541 aaacacctca atggggcagc attcagcaaa gacctccaat cagcgaaaat agcagatctg
     3601 tagctgctac tatccccagg cgcaataccc tccctgcaat ggacaacagc cgactccatc
     3661 gagcttcttc ctgtgagacc tacgagtacc cggcacgagg cagtggggaa agtgccagct
     3721 ggtctgctga acctccagga aagactgccg taggtcgatc aaatagtgcc agctctgatg
     3781 acaactacgt gcccatgaac ccaggttctt ctaccctgct ggctatggaa cgaccagggg
     3841 acaactccca gagtgtctac atccccatga gcccaggacc ccatcacttt gacccacttg
     3901 gctacccgtc cacagccctt cctattcaca gaggccccag ccgaggaagt gaaatccagc
     3961 cacccccggt caaccgaaac ctcaagcctg acagaaaagc aaagccaaca ccccttgacc
     4021 tgagaaacaa cactgtcatc gatgagctgc ccttcaagtc acctgtcacc aagtcttggt
     4081 ccaggatcaa cagccacacc tttaactcca gttcctccca gtactgccgt cctatctcca
     4141 cacagagcat caccagcaca gactcgggag acagtgagga gaactatgtc cctatgcaaa
     4201 acccagtatc tgcatcccct gttcccagtg gcactaacag cccagctcca aagaagagta
     4261 ctggcagtgt ggattatctc gccctggact tccagccggg ctccccaagc cctcaccgca
     4321 agccatccac atcatctgtc acatcagatg agaaggtaga ctatgtccaa gtggataaag
     4381 agaagaccca ggccctgcag aacaccatgc aggagtggac agatgtgcgg cagtcctccg
     4441 aaccttccaa gggtgccaag ctgtagggtt aacgaattcc gccccccccc cctaacgtta
     4501 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     4561 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     4621 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     4681 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     4741 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     4801 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     4861 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     4921 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     4981 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     5041 gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     5101 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc
     5161 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     5221 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     5281 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     5341 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     5401 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     5461 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     5521 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
     5581 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     5641 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     5701 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     5761 gacgagctgt acaagtaatg aattaattaa gaattatcaa gcttatcgat accgtcgacc
     5821 tgcagccaag cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     5881 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     5941 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     6001 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6061 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     6121 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6181 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6241 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata
     6301 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     6361 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
     6421 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
     6481 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
     6541 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6601 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6661 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6721 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6781 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6841 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6901 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6961 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7021 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7081 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7141 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7201 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7261 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7321 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7381 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7441 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7501 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7561 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7621 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7681 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7741 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7801 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7861 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7921 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7981 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8041 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8101 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8161 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8221 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8281 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8341 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8401 gctgttgaga tccagttcga tgtaacccac tcgtgcac
//
