LOCUS       sequence-262411-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSIREN-RetroQ vector containing shRNA sequence to Egr3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  AUTHORS   Salotti J, Sakchaisri K, Tourtellotte WG, Johnson PF
  TITLE     An Arf-Egr-C/EBPbeta pathway linked to ras-induced senescence and 
            cancer.
  JOURNAL   Mol Cell Biol. 2015 Mar;35(5):866-83. doi: 10.1128/MCB.01489-14. 
            Epub 2014 Dec 22.
  PUBMED    25535333
REFERENCE   2  (bases 1 to 6503)
  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"; doi: 
            "10.1128/MCB.01489-14"; journalName: "Mol Cell Biol"; date: 
            "2015-03"; volume: "35"; issue: "5"; pages: "866-83"
COMMENT     Sequence Label: pSirenRetroQ(puro)-shEgr3
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             586..761
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    824..1023
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1224..1640
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1570..1592
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1608..1624
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1664..1912
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1664..1684
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1835..1854
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        1982..2481
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2017..2040)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2414..2433
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2502..3101
                     /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(2502..2521)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2998..3018
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3282..3707
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(3810..3832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3968..3988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3973..4302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4153..4288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4324..4343
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4424..4441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4595..5183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4675..4694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5354..6145,6146..6214))
                     /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     5977..5996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6215..6319)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6387..6405
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata aaagagccca
      601 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
      661 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
      721 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
      781 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
      841 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
      901 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
      961 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     1021 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     1081 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     1141 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     1201 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     1261 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     1321 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     1381 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     1441 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     1501 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     1561 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     1621 tcactccttc tctaggcgcc ggaattgaag atctgggcag gaagagggcc tatttcccat
     1681 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
     1741 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     1801 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     1861 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaggat ccaatcaatt
     1921 gcctgacaat cttcaagaga gattgtcagg caattgattt tttttacgcg tgaattctac
     1981 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
     2041 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
     2101 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
     2161 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
     2221 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
     2281 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
     2341 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
     2401 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
     2461 tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca
     2521 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     2581 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     2641 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     2701 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     2761 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     2821 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     2881 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     2941 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     3001 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     3061 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc
     3121 agcgcccgac cgaaaggagc gcacgacccc atgcatcggc gtctcgagat atcagtggtc
     3181 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     3241 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     3301 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     3361 agaatagaga agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac
     3421 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     3481 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     3541 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     3601 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     3661 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
     3721 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc
     3781 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3841 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3901 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc
     3961 cggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4021 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4261 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact
     4321 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     4381 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     4441 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4501 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5641 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     5701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5941 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     6001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6361 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     6421 aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
     6481 catgtccaac attaccgcca tgt
//
