LOCUS       sequence-178015-        4411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of control shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4411)
  AUTHORS   Oloris SC, Frazer-Abel AA, Jubala CM, Fosmire SP, Helm KM, Robinson 
            SR, Korpela DM, Duckett MM, Baksh S, Modiano JF
  TITLE     Nicotine-mediated signals modulate cell death and survival of T 
            lymphocytes.
  JOURNAL   Toxicol Appl Pharmacol. 2010 Feb 1;242(3):299-309. doi: 
            10.1016/j.taap.2009.10.020. Epub 2009 Nov 4.
  PUBMED    19896492
REFERENCE   2  (bases 1 to 4411)
  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.taap.2009.10.020"; journalName: "Toxicol Appl Pharmacol";
            date: "2010-02-1- 1"; volume: "242"; issue: "3"; pages: "299-309"
COMMENT     Sequence Label: alpha7-nAChR(scramble control)
FEATURES             Location/Qualifiers
     source          1..4411
                     /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(292..308)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(293..309)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(337..355)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(376..392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(376..392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(389..411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    400..416
                     /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(424..430,431..448,449..454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    469..490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(607..624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(778..1366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(858..877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1537..2328,2329..2397))
                     /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     2160..2179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2398..2502)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2528..2983)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2665..2686)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2877..2896
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3110..3132
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3124..3141
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3125..3141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3148..3167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3148..3166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3199..3215
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3199..3215
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(3278..3877)
                     /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(3361..3381)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3858..3877
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3902..4401)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3950..3969)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     4343..4366
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse 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 cccgctcctc atatatcatc
      241 aatattcaag agatattgat gatatatgag gagctttttg gaaaagctta tcgataccgt
      301 cgacctcgag ggggggcccg gtaccagctt ttgttccctt tagtgagggt taatttcgag
      361 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
      421 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
      481 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      541 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
      601 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      661 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      721 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      781 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      841 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      901 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      961 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1021 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1081 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1141 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1201 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1261 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1321 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1381 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1441 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1501 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1561 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1621 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1681 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1741 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1801 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1861 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1921 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1981 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2041 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2101 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2161 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2221 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2281 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2341 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2401 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2461 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2521 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2581 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2641 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2701 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2761 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2821 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2881 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2941 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     3001 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3061 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3121 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attggagctc
     3181 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcatgg ggtcgtgcgc
     3241 tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca
     3301 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     3361 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc
     3421 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     3481 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     3541 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     3601 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     3661 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     3721 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     3781 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     3841 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt aagctagctt gggctgcagg
     3901 tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc ttttgaagcg
     3961 tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc
     4021 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa
     4081 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca
     4141 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct
     4201 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac
     4261 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc
     4321 cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca gactgccttg
     4381 ggaaaagcgc ctcccctacc cggtagaatt c
//
