LOCUS       sequence-217045-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector designed to knock down LARP7..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  AUTHORS   Shelton SB, Shah NM, Abell NS, Devanathan SK, Mercado M, Xhemalce B
  TITLE     Crosstalk between the RNA Methylation and Histone-Binding Activities
            of MePCE Regulates P-TEFb Activation on Chromatin.
  JOURNAL   Cell Rep. 2018 Feb 6;22(6):1374-1383. doi: 
            10.1016/j.celrep.2018.01.028.
  PUBMED    29425494
REFERENCE   2  (bases 1 to 6372)
  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: 
            "10.1016/j.celrep.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-6- 6"; volume: "22"; issue: "6"; pages: "1374-1383"
COMMENT     Sequence Label: pSUPER.retro.puro-shLARP7
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..222
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     143..162
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(291..307)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(292..308)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(326..742)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(342..358)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(374..396)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    809..1150
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             1214..1730
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1930..1949)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2077..2099)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2293..2312)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2412..2434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2472..2490)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2558..2662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2663..2731,2732..3523)
                     /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     complement(2881..2900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3694..4282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4183..4202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4436..4453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4570..4591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4606..4611,4612..4629,4630..4636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4644..4660
                     /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)."
     CDS             complement(5246..5845)
                     /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(5329..5349)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5826..5845
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5870..6369)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5918..5937)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6311..6334
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 agaattcgaa cgctgacgtc atcaacccgc tccaaggaat cgcgggccca gtgtcactag
       61 gcgggaacac ccagcgcgcg tgcgccctgg caggaagatg gctgtgaggg acaggggagt
      121 ggcgccctgc aatatttgca tgtcgctatg tgttctggga aatcaccata aacgtgaaat
      181 gtctttggat ttgggaatct tataagttct gtatgagacc acagatcccc aagttaatca
      241 ccaaagctgt tcaagagaca gctttggtga ttaacttttt ttaagcttat cgataccgtc
      301 gacctcgaga gatcgatcta attccggcgc ctagagaagg agtgagggct ggataaaggg
      361 aggatcgagg cggggtcgaa cgaggaggtt caagggggag agacggggcg gatggaggaa
      421 gaggaggcgg aggcttaggg tgtacaaagg gcttgaccca gggagggggg tcaaaagcca
      481 aggcttccca ggtcacgatg taggggacct ggtctgggtg tccatgcggg ccaggtgaaa
      541 agaccttgat cttaacctgg gtgatgaggt ctcggttaaa ggtgccgtct cgcggccatc
      601 cgacgttaaa ggttggccat tctgcagagc agaaggtaac ccaacgtctc ttcttgacat
      661 ctaccgactg gttgtgagcg atccgctcga catctttcca gtgacctaag gtcaaactta
      721 agggagtggt aacagtctgg ccctaatttt cagacaaata cagaaacaca gtcagacaga
      781 gacaacacag aacgatgctg cagcgctgca gcagacaaga cgcgcggctt cggttccaaa
      841 ccgaaagcaa aaattcagac ggaggcggga actgttttag gttctcgtct cctaccagaa
      901 ccacatatcc tgacggggtc ggattccaca tcgactccct tcctcaggtc gggccacaaa
      961 aacggccccc aaagtccctg ggacgtctcc cagggttgcg gccgggtgtt cagaactcgt
     1021 cagttccacc acgggtccgc cagatacaga gctagttagc taactagtac agacgcaggc
     1081 gcaaacatta gatgccggca caaacacgca caaagacaga gacagacacg aaacgaccgc
     1141 tggccagctt acctcccggc gggggggtcg gtggtccctg ggcaggggtc tccaaatctc
     1201 ggtggaacct ccaaatgaaa gacccccgag gtgggcagtc aatcaatctg aggagaccct
     1261 cccaaggatc agcgagtcca cgattcggat gcaaacagca agaggcttta ttgggaatac
     1321 gggtacccgg gcgacgcagt ctatcggagg actggcgcgc cgagtgaggg gttgtgggct
     1381 cttttattga gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt
     1441 tagttcaaat aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg
     1501 gttctctaga aactgctgag ggcgggaccg catctgggga ccatctgttc ttggccctga
     1561 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattctg ctgtctctct
     1621 gttcctaacc ttgatctgaa cttctctatt ctcagttatg tattttccat gccttgcaaa
     1681 atggcgttac ttaagctagc ttgccaaacc tacaggtggg gtctttcatt cccccctttt
     1741 tctggagact aaataaaatc ttttatttta tctatggctc gtactctata ggcttcagct
     1801 ggtgatattg ttgagtcaaa actagagcct ggaccactga tatcctgtct ttaacaaatt
     1861 ggactaatcg cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1921 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1981 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     2041 gcgccatctc cttgcatgca ccattccttg cgccgtcgtt ttacaacgtc gtgactggga
     2101 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     2161 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2221 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     2281 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2341 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2401 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2461 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2521 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2581 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2641 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2881 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3181 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3421 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3541 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3841 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4321 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4381 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4441 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4501 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4561 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4621 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacaatt ctccccagct
     4681 ctttctgcgc tgtccataat gaactatttc aggagtattg gagtatgtgg attcaaaaca
     4741 gtggctaatt gagtcaggat tacttaatat tcgactccta ccctcagaat gttgttctcc
     4801 aacttcaaga aactgttacc catgaaagac ccccgctgac gggtagtcaa tcactcagag
     4861 gagaccctcc caaggaacag cgagaccaca agtcggatgc aactgcaaga gggtttattg
     4921 gatacacggg tacccgggcg acgcagtcta tcggaggact ggcgcgccga gtgaggggtt
     4981 gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga gaagcgaact
     5041 gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac cctggaaaca
     5101 tctgatggtt ctctagcttg ccaaacctac aggtggggtc tttcattccc ccctttttct
     5161 ggagactaaa taaaatcttt tattttatcg atgcatgggg tcgtgcgctc ctttcggtcg
     5221 ggcgctgcgg gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg caccaggtgc
     5281 gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg
     5341 ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct
     5401 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc
     5461 cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca
     5521 tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg
     5581 cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt
     5641 cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca
     5701 gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt
     5761 cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc
     5821 gcaccgtggg cttgtactcg gtcatggtaa gctagcttgg gctgcaggtc gaaaggcccg
     5881 gagatgagga agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg
     5941 ggcctccgga ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc
     6001 cggacccacc ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc
     6061 cgctgcccca aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac
     6121 tagtgagacg tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg
     6181 ggaacttcct gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag
     6241 ccggttggcg cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc
     6301 gccaagtgcc cagcggggct gctaaagcgc atgctccaga ctgccttggg aaaagcgcct
     6361 cccctacccg gt
//
