LOCUS       sequence-171278-        6809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control hairpin sequence used in gene knockdown experiements.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  AUTHORS   Swartz KL, Wood SN, Murthy T, Ramirez O, Qin G, Pillai MM, Rao S, 
            Minella AC
  TITLE     E2F-2 promotes nuclear condensation and enucleation of terminally 
            differentiated erythroblasts.
  JOURNAL   Mol Cell Biol. 2016 Oct 17. pii: MCB.00274-16.
  PUBMED    27795297
REFERENCE   2  (bases 1 to 6809)
  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: "Mol Cell 
            Biol. 2016 Oct 17. pii: MCB.00274-16."
COMMENT     Sequence Label: Banshee-Sh-Ctrl
FEATURES             Location/Qualifiers
     source          1..6809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(1178..1194)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1178..1194)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1231..1249)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1270..1286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1270..1286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1283..1305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1294..1310
                     /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)."
     promoter        complement(join(1318..1324,1325..1342,1343..1348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1363..1384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1501..1518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1672..2260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1752..1771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2431..3222,3223..3291))
                     /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     3054..3073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3292..3396)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3422..3877)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3559..3580)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3771..3790
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4004..4026
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4018..4035
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4019..4035
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4042..4061
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4042..4060
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             4523..4698
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    4762..5103
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             5170..5586
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5516..5538
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5554..5570
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      5632..5641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5638..5640,5641..5643,5644..6357)
                     /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(5638..5640,5641..5643,5644..6354)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5683..5704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5944..5963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6291..6312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6468..6487
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacagatc ttatgtcaag ttgtatagtt
      241 attcaagaga taactataca acttgacata tttttaagct tggatcaagg tcagaaacag
      301 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
      361 agggccaaga acagttggaa cagcagaata tgggccaaac aggatatctg tggtaagcag
      421 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
      481 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      541 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      601 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
      661 cgcccgggta cccgtgttct caataaaccc tcttgcagtt gcatccgact cgtggtctcg
      721 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcagt
      781 ttctcccacc tacacaggtc tcactaacat tcctgatgtg ccgcagggac tccgtcagcc
      841 cggtttttgt ttataataaa atgcaagaac agtgttccct tcaagccaga ctacatcctg
      901 actctcggct ttataaaaga atgttgaagg gctctgtgga ctatctgcca cacgactttt
      961 aagattttta tgcctcctgg atgagggatt tagtcaatct atcctcgtct attttgctgg
     1021 cttctccgta ttttaaattt ctagtttgca ctcccttcct gagagcacgg cgattgcaga
     1081 gtagttaata ctctgagggc aggcttctgt gaaaaggttg cctgggctca gtgtgagatt
     1141 ttgccataaa aaggggtcct gcccctgtgt acagacagat ccactagttc tagagcggcc
     1201 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattt cgagcttggc
     1261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1321 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2101 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3421 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3481 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3541 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3601 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3661 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3721 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3781 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3841 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     3901 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3961 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     4021 aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg taccgggccc
     4081 ccccgccgcg gcccttccgt ttctttgctt ttgaaagacc ccacccgtag gtggcaagct
     4141 agcttaagta acgccacttt gcaaggcatg gaaaaataca taactgagaa tagaaaagtt
     4201 cagatcaagg tcaggaacaa agaaacagct gaataccaaa caggatatct gtggtaagcg
     4261 gttcctgccc cggctcaggg ccaagaacag atgagacagc tgagtgatgg gccaaacagg
     4321 atatctgtgg taagcagttc ctgccccggc tcggggccaa gaacagatgg tccccagatg
     4381 cggtccagcc ctcagcagtt tctagtgaat catcagatgt ttccagggtg ccccaaggac
     4441 ctgaaaatga ccctgtacct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     4501 gcgcgcttcc gctctccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     4561 tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg
     4621 catccgaatc gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca
     4681 cctcgggggt ctttcatttg gaggttccac cgagatttgg agacccctgc ccagggacca
     4741 ccgacccccc cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt
     4801 gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt agctaactag
     4861 ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct
     4921 gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg
     4981 atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag
     5041 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
     5101 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     5161 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
     5221 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
     5281 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
     5341 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
     5401 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
     5461 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
     5521 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
     5581 ggcgccggaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg
     5641 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5701 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     5761 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     5821 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     5881 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     5941 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     6001 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     6061 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     6121 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     6181 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     6241 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     6301 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     6361 ggcccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6421 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6481 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6541 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6601 gggactctgg ggttcggtag tcgacggatc ctctagacgc gtctcgagtc gacggtatcg
     6661 attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca
     6721 atatcacagc tgaagcctat agagtacgag ccatagatag aataaaagat tttatttagt
     6781 ctccagaaaa aggggggaat gaaagatcc
//
