LOCUS       sequence-254627-        6919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible mammalian retroviral expression of HuR (S221D)
            with FLAG and turboGFP tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6919)
  TITLE     Mammalian retroviral expression plasmids based on pBABE family
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6919)
  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            "Unpublished"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: pBABE.puro/TO/Flag/tGFP.HuR.S221D
FEATURES             Location/Qualifiers
     source          1..6919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1361..1379
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1389..1413
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1503..1512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1512..1535
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1542..2237
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     primer_bind     complement(3242..3262)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3247..3576
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3427..3562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3586..4185
                     /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(3586..3605)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4082..4102
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4406..4875
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5005..5022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5176..5764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5256..5275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5924..6715,6716..6784))
                     /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     6547..6566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6785..6889)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc tccctatcag tgatagagat
     1381 cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     1441 gacctccata gaagacaccg ggaccgatcc agcctccgga ctctagcgtt taaacttaag
     1501 ctgccaccat ggactacaag gacgacgatg acaagaagct tatggagagc gacgagagcg
     1561 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg
     1621 agctggtggg cggcggagag ggcacccccg agcagggccg catgaccaac aagatgaaga
     1681 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct
     1741 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca
     1801 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga
     1861 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg atgggcaccg
     1921 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg
     1981 agcacctgca ccccatgggc gataacgatc tggatggcag cttcacccgc accttcagcc
     2041 tgcgcgacgg cggctactac agctccgtgg tggacagcca catgcacttc aagagcgcca
     2101 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagg
     2161 atcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag accccggatg
     2221 cagatgccgg tgaagaaaga gttggtacca tgtctaatgg ttatgaagac cacatggccg
     2281 aagactgcag gggtgacatc gggagaacga atttgatcgt caactacctc cctcagaaca
     2341 tgacccagga tgagttacga agcctgttca gcagcattgg tgaagttgaa tctgcaaaac
     2401 ttattcggga taaagtagca ggacacagct tgggctatgg ctttgtgaac tacgtgaccg
     2461 cgaaggatgc agagagagcg atcaacacgc tgaacggctt gaggctccag tcaaaaacca
     2521 ttaaggtgtc gtatgctcgc ccgagctcag aggtgatcaa agacgccaac ttgtacatca
     2581 gcgggctccc gcggaccatg acccagaagg acgtagaaga catgttctct cggtttgggc
     2641 ggatcatcaa ctcgcgggtc ctcgtggatc agactacagg tttgtccaga ggggttgcgt
     2701 ttatccggtt tgacaaacgg tcggaggcag aagaggcaat taccagtttc aatggtcata
     2761 aacccccagg ttcctctgag cccatcacag tgaagtttgc agccaacccc aaccagaaca
     2821 aaaacgtggc actcctctcg cagctgtacc actcgccagc gcgacggttc ggaggccccg
     2881 ttcaccacca ggcgcagaga ttcaggttcg accccatggg cgtcgatcac atgagcgggc
     2941 tctctggcgt caacgtgtca ggaaacgcct cctccggctg gtgcattttc atctacaacc
     3001 tggggcagga tgccgacgag gggatcctct ggcagatgtt tgggccgttt ggtgccgtca
     3061 ccaatgtgaa agtgatccgc gacttcaaca ccaacaagtg caaagggttt ggctttgtga
     3121 ccatgacaaa ctatgaagaa gccgcgatgg ccatagccag cctgaacggc taccgcctgg
     3181 gggacaaaat cttacaggtt tccttcaaaa ccaacaagtc ccacaaataa ctcgaccctg
     3241 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3301 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3361 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3421 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3481 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3541 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag
     3601 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3661 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3721 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3781 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3841 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3901 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3961 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4021 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4081 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4141 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     4201 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgataaaata aaagatttta
     4261 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     4321 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     4381 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     4441 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     4501 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4561 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4621 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4681 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     4741 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4801 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4861 agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     4921 ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc
     5881 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5941 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6001 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6061 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6121 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6181 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6241 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6301 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6361 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6421 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6481 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6541 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6601 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6661 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6721 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6781 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6841 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6901 gaaaagtgcc acctgcagc
//
