LOCUS       sequence-190135-        7610 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express GATA3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7610)
  AUTHORS   Sun J, He H, Pillai S, Xiong Y, Challa S, Xu L, Chellappan S, Yang S
  TITLE     GATA3 transcription factor abrogates Smad4 transcription 
            factor-mediated fascin overexpression, invadopodium formation, and 
            breast cancer cell invasion.
  JOURNAL   J Biol Chem. 2013 Dec 27;288(52):36971-82. doi: 
            10.1074/jbc.M113.506535. Epub 2013 Nov 14.
  PUBMED    24235142
REFERENCE   2  (bases 1 to 7610)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M113.506535"; journalName: "J Biol Chem"; date: 
            "2013-12-27- 27"; volume: "288"; issue: "52"; pages: "36971-82"
COMMENT     Sequence Label: pLPCX2-HA-GATA3 FL
FEATURES             Location/Qualifiers
     source          1..7610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             561..587
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             2011..2604
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2707..2729)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    2857..2890
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2950..2983
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3003..3022
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3103..3120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3274..3862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3354..3373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4033..4824,4825..4893))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4656..4675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4894..4998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5066..5084
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5686..5861
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5924..6123
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6324..6740
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6670..6692
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6708..6724
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6837..7436
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6837..6856)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7333..7353
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctatggac tacccttatg atgtgccgga ttatgccgag gtgactgcgg
      601 accagccgcg ctgggtgagc caccatcacc ccgcggtcct caacggtcag cacccagaca
      661 cgcaccaccc gggcctcggc cattcgtaca tggaagctca gtatccgctg acggaagagg
      721 tggacgtact ttttaacatc gatggtcaag gcaaccacgt cccgtcctac tacggaaact
      781 ccgtcagggc tacggtgcag aggtatcctc cgacccacca cgggagccag gtatgccgcc
      841 cgcctctgct gcacggatct ctgccctggc tggatggcgg caaagccctg agcagccacc
      901 acaccgcctc gccctggaac ctcagcccct tctccaagac gtccatccac cacggctctc
      961 cggggcctct gtccgtttac cctccggctt catcctcttc tctggcggcc ggccactcca
     1021 gtcctcatct cttcaccttc ccgcccaccc cgccgaaaga cgtctcccca gacccgtcgc
     1081 tgtccacccc gggatccgcc gggtcggcca ggcaagatga gaaagagtgc ctcaagtatc
     1141 aggtgcagct gccagatagc atgaagctgg agacgtctca ctctcgaggc agcatgacca
     1201 ccctgggtgg ggcctcatcc tcagcccacc accccattac cacctatccg ccctatgtgc
     1261 ccgagtacag ctctggactc ttcccaccca gcagcctgct gggaggatcc cctaccgggt
     1321 tcggatgtaa gtcgaggccc aaggcacgat ccagcacaga aggcagggag tgtgtgaact
     1381 gcggggcaac ctctacccca ctgtggcggc gagatggtac cgggcactac ctttgcaatg
     1441 cctgcggact ctaccataaa atgaatgggc agaaccggcc ccttatcaag cccaagcgaa
     1501 ggctgtcggc agcaaggaga gcagggacat cctgcgcgaa ctgtcagacc accaccacca
     1561 ccctctggag gaggaacgct aatggggacc cggtctgcaa tgcctgtggg ctgtactaca
     1621 agcttcataa tattaacaga cccctgacta tgaagaaaga aggcatccag acccgaaacc
     1681 ggaagatgtc tagcaaatcg aaaaagtgca aaaaggtgca tgacgcgctg gaggacttcc
     1741 ccaagagcag ctccttcaac ccggccgctc tctccagaca catgtcatcc ctgagccaca
     1801 tctctccctt cagccactcc agccacatgc tgaccacacc gacgcccatg catccgccct
     1861 ccggcctctc cttcggacct caccaccctt ccagcatggt caccgccatg ggtagaattc
     1921 gttaggccat taaggcctgt cgacaagcgg ccgcctcggc caaacatcga taaaataaaa
     1981 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     2041 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     2101 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     2161 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     2221 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     2281 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     2341 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     2401 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     2461 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     2521 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     2581 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
     2641 ggaccaccga cccaccaccg ggaggtaagc tggctgcctc gcgcgtttcg gtgatgacgg
     2701 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2761 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     2821 catgacccag tcacgtagcg atagcggagt gtactgataa cttcgtataa tgtatgctat
     2881 acgaagttat taggtctgaa gaggagttta cgtccagcca agctagggcc gcgatccgga
     2941 acccttaata taacttcgta taatgtatgc tatacgaagt tatcagtact ggcttaacta
     3001 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     3061 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     3121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3181 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3241 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4141 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4321 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     4381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4621 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     4681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4861 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4981 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5041 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     5101 attgctagca attgctagca attgctagca attcatacca gatcaccgaa aactgtcctc
     5161 caaatgtgtc cccctcacac tcccaaattc gcgggcttct gcctcttaga ccactctacc
     5221 ctattcccca cactcaccgg agccaaagcc gcggcccttc cgtttctttg cttttgaaag
     5281 accccacccg taggtggcaa gctagcttaa gtaacgccac tttgcaaggc atggaaaaat
     5341 acataactga gaatagaaaa gttcagatca aggtcaggaa caaagaaaca gctgaatacc
     5401 aaacaggata tctgtggtaa gcggttcctg ccccggctca gggccaagaa cagatgagac
     5461 agctgagtga tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcggggc
     5521 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagtg aatcatcaga
     5581 tgtttccagg gtgccccaag gacctgaaaa tgaccctgta ccttatttga actaaccaat
     5641 cagttcgctt ctcgcttctg ttcgcgcgct tccgctctcc gagctcaata aaagagccca
     5701 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
     5761 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
     5821 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
     5881 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
     5941 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
     6001 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
     6061 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     6121 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     6181 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     6241 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     6301 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     6361 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     6421 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     6481 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     6541 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     6601 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     6661 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     6721 tcactccttc tctaggcgcc ggaatgatcc cccgggctgc aggtcggccg ccacgaccgg
     6781 tgccgccacc atcccctgac ccacgcccct gacccctcac aaggagacga ccttccatga
     6841 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     6901 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     6961 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     7021 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     7081 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     7141 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     7201 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     7261 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     7321 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     7381 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc
     7441 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc tccgaccgaa
     7501 gccacccggg gcggccccgc cgaccccgca cccgcccccg aggcccaccg actctagtaa
     7561 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
