LOCUS       sequence-166278-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GATA3 fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7319)
  TITLE     Engel Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7319)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCMV-GATA3gfp
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             913..921
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             join(3272..3274,3275..3277,3278..3991)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3272..3274,3275..3277,3278..3988)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3317..3338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3578..3597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3925..3946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4114..4235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4151..4170)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4205..4224
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4242..4697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4591..4610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4724..4828
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4830..5187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4830..5021
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4853..4873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5038..5173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5222..6016
                     /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             5222..6016
                     /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(5276..5295)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5886..5905
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6204..6223)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6248..6295
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6624..7212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7113..7132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ccctcctgcc tgtcccctct
      661 gccccaagcc cgggcgggcc tcgcccaccc tgcccgcggc ccctctgccc taggcggccc
      721 ttggcggcgc ccctttccgg tcagtagagg ggcgggagga ggggcggcgg tgtgccgggg
      781 tgggtggggg ggatgggatg ggggtggagg tgaagtcctg gagcgggttt gggttgcagt
      841 ttccttgtgc tgaggatcgt gtcccctcct gccagcgcca ggctcctccc cctcggcgcg
      901 gatgacacta gaacctcctt aagttgcgtc gcgccacagc tgtctgcgaa cactgagctg
      961 cctggcgccg tcttgatagt ttcagaaaga atgcattccc tgtaaaaaaa aaatactgag
     1021 agagggagag gagaaagaga gagagactga gagagcgaga catagagagc tacgcaatct
     1081 gaccgggcag gtcacacgcc tcctcctcct cctctacgct ccttgctact caggttggta
     1141 ttgtgacttg tgactttttt ttttttttta agtttgatat cactccctcc tcttggaggg
     1201 actgtttgtg tgtgtgtgtt tgggggtttg tttgtttggt tttttgtaaa ctaaaagctc
     1261 agctttctcc ttttggagga tttgtagtgg ttggaaagat cctggggccg gtgtcctcca
     1321 gggaggactt gtccccaagt tttaattgct tagtcagtcc tgggctcctg cgagcctggc
     1381 tgtcggaggt gtgctgcgga gcttggcttc tttccctccg ggtcttcgcg tgcgtgtgca
     1441 cgtcccgggt gagtcggccc tgcaggaagc ttgcgaagac ctagtgccct ccagaatttc
     1501 aggtgttaaa agttcttaca cctcattcag gtctctcttt ctctcccttg caggtgatcg
     1561 gaagagcaac cgtctctgag cgccaaggaa tcagtgtgca gtgtggtcac actcggattc
     1621 ctctctccct cctttttttt tttttttttt tgaccccttt attcctccgt gtctgctttt
     1681 tttttggggg gggggatcgc cctcattctt ttctttttct tctttccctt cctttccttt
     1741 gctaaactat cccgcaaaga tttttctttc ctccctaaac cctccttttt gctctccttt
     1801 tctataccct taactgcaaa caaaccatta aacgacccct ctcctgggcc tccgacggca
     1861 ggagtccgcg gacctcccag gccgacagcc ctccctctac ccgcgagggt tccgggccgg
     1921 gcgagagggc gcgagcacag ccgaggccat ggaggtgact gcggaccagc cgcgctgggt
     1981 gagccaccat caccccgcgg tcctcaacgg tcagcaccca gacacgcacc acccgggcct
     2041 cggccattcg tacatggaag ctcagtatcc gctgacggaa gaggtggacg tactttttaa
     2101 catcgatggt caaggcaacc acgtcccgtc ctactacgga aactccgtca gggctacggt
     2161 gcagaggtat cctccgaccc accacgggag ccaggtatgc cgcccgcctc tgctgcacgg
     2221 atctctgccc tggctggatg gcggcaaagc cctgagcagc caccacaccg cctcgccctg
     2281 gaacctcagc cccttctcca agacgtccat ccaccacggc tctccggggc ctctgtccgt
     2341 ttaccctccg gcttcatcct cttctctggc ggccggccac tccagtcctc atctcttcac
     2401 cttcccgccc accccgccga aagacgtctc cccagacccg tcgctgtcca ccccgggatc
     2461 cgccgggtcg gccaggcaag atgagaaaga gtgcctcaag tatcaggtgc agctgccaga
     2521 tagcatgaag ctggagacgt ctcactctcg aggcagcatg accaccctgg gtggggcctc
     2581 atcctcagcc caccacccca ttaccaccta tccgccctat gtgcccgagt acagctctgg
     2641 actcttccca cccagcagcc tgctgggagg atcccctacc gggttcggat gtaagtcgag
     2701 gcccaaggca cgatccagca cagaaggcag ggagtgtgtg aactgcgggg caacctctac
     2761 cccactgtgg cggcgagatg gtaccgggca ctacctttgc aatgcctgcg gactctacca
     2821 taaaatgaat gggcagaacc ggccccttat caagcccaag cgaaggctgt cggcagcaag
     2881 gagagcaggg acatcctgcg cgaactgtca gaccaccacc accaccctct ggaggaggaa
     2941 cgctaatggg gacccggtct gcaatgcctg tgggctgtac tacaagcttc ataatattaa
     3001 cagacccctg actatgaaga aagaaggcat ccagacccga aaccggaaga tgtctagcaa
     3061 atcgaaaaag tgcaaaaagg tgcatgacgc gctggaggac ttccccaaga gcagctcctt
     3121 caacccggcc gctctctcca gacacatgtc atccctgagc cacatctctc ccttcagcca
     3181 ctccagccac atgctgacca caccgacgcc catgcatccg ccctccggcc tctccttcgg
     3241 acctcaccac ccttccagca tggtcaccgc catggtgagc aagggcgagg agctgttcac
     3301 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3361 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3421 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3481 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3541 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3601 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3661 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3721 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3781 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3841 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     3901 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3961 cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
     4021 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4081 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4141 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4201 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     4261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4681 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4801 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4861 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4921 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4981 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5041 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5101 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5161 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5221 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5281 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5341 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5401 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5461 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5521 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5581 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5641 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5701 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5761 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5821 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5881 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5941 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6001 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6061 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6121 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6181 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6241 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6301 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6361 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6421 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6481 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6541 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7081 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7141 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7201 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7261 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgc
//
