LOCUS       sequence-239053-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  AUTHORS   Kishi Y, Fujii Y, Hirabayashi Y, Gotoh Y
  TITLE     HMGA regulates the global chromatin state and neurogenic potential 
            in neocortical precursor cells.
  JOURNAL   Nat Neurosci. 2012 Aug;15(8):1127-33. doi: 10.1038/nn.3165.
  PUBMED    22797695
REFERENCE   2  (bases 1 to 6563)
  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: "10.1038/nn"; 
            journalName: "Nat Neurosci"; date: "2012-08"; volume: "15"; issue: 
            "8"; pages: "1127-33"
COMMENT     Sequence Label: pSiren_EGFP-shLuc
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..568
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(104..127)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     501..520
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(589..591,592..594,595..1308)
                     /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(589..591,592..594,595..1305)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(634..655)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(895..914)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1242..1263
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1430..1855
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(1958..1980)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2116..2136)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2121..2450
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2301..2436
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2472..2491
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2572..2589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2743..3331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2823..2842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3502..4293,4294..4362))
                     /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     4125..4144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4363..4467)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4535..4553
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4653..5032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5033..5236
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5187..5207
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5237..5412
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5475..5674
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5875..6291
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6221..6243
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6259..6275
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        6315..6563
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6315..6335
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6486..6505
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gtgcgttgct agaccaactt caagagagtt ggtactagca acgcactttt ttgctagcga
       61 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
      121 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
      181 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
      241 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
      301 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
      361 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
      421 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
      481 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
      541 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat ggtgagcaag
      601 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      661 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      721 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      781 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      841 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      901 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      961 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1021 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1081 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1141 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1201 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1261 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag ctcgagatat
     1321 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     1381 agccatagat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     1441 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     1501 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gggtcgaccc
     1561 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     1621 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     1681 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     1741 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     1801 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttgggg
     1861 gctcgtccgg gatcgggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
     1921 ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     1981 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2041 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     2101 tgtagatccg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2161 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2221 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2281 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2341 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     2401 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttactg
     2461 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     2521 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     2581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3181 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3781 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     3841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     4141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4501 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4561 tcttcaagaa ttagcttggc cattgcatac gttgtatcca tatcataata tgtacattta
     4621 tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata
     4681 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4741 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4801 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4861 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4921 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4981 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     5041 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     5101 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     5161 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     5221 ctatataagc agagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcttcc
     5281 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
     5341 aatcgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccacgacgg
     5401 gggtctttca tttgggggct cgtccgggat ttggagaccc ctgcccaggg accaccgacc
     5461 caccaccggg aggtaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct
     5521 atgtttgatg ttatgcgcct gcgtctgtac tagttagcta actagctctg tatctggcgg
     5581 acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg
     5641 gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc
     5701 ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct
     5761 gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc
     5821 atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag
     5881 actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct
     5941 cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg
     6001 ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag
     6061 gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc
     6121 gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct
     6181 aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg
     6241 accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggaattgaa
     6301 gatctgggca ggaagagggc ctatttccca tgattccttc atatttgcat atacgataca
     6361 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     6421 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     6481 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     6541 tcttgtggaa aggacgagga tcc
//
