LOCUS       sequence-424965-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Wang L, Brugge JS, Janes KA
  TITLE     Intersection of FOXO- and RUNX1-mediated gene expression programs in
            single breast epithelial cells during morphogenesis and tumor 
            progression.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Oct 4;108(40):E803-12. doi: 
            10.1073/pnas.1103423108. Epub 2011 Aug 22.
  PUBMED    21873240
REFERENCE   2  (bases 1 to 7083)
  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: 
            "10.1073/pnas.1103423108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-10-4- 4"; volume: "108"; issue: "40"; pages: "E803-12"
COMMENT     Sequence Label: pLKO.1 shRUNX1 puro
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..34)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(188..776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(268..287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(947..1738,1739..1807))
                     /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     1570..1589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1808..1912)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1938..2393)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2075..2096)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2287..2306
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2520..2542
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2534..2551
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2535..2551
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2561..2580
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2561..2579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2600..2735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2775..2896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2786..2805)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2840..2859
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2968..3201
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(3329..3928)
                     /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(3412..3432)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3909..3928
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3950..4460)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4013..4031)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    4509..4626
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(4722..4970)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4780..4799)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4950..4970)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(5268..5312)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5497..5730
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6223..6348
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6395..6575
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6576..6802)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(6830..6850)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6830..6848)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6869..6885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6869..6885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6882..6904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6893..6909
                     /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(6917..6923,6924..6941,6942..6947))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6962..6983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
       61 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      601 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      901 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      961 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1021 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1081 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1141 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1201 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1261 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1321 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1381 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1441 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1501 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1561 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1621 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1681 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1741 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1801 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1861 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1921 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1981 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2041 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2101 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2161 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2221 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2281 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2341 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     2401 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2461 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2521 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     2581 cgaattgggt acgtccctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     2641 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     2701 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggatagct
     2761 agagccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2821 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2881 ctgcaataaa caagttcctc tcactctctg atattcattt ctttgcaagt tataaatact
     2941 gaataataag atgacatgaa ctactactgc tagagatttt ccacactgac taaaagggtc
     3001 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     3061 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     3121 ctcagatctg gtctaaccag agagacccag tacaagcaaa aagcagatct tgtcttcgtt
     3181 gggagtgaat tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag
     3241 ctgccttgta agtcattggt cttaaaggta ccgatgcatg gggtcgtgcg ctcctttcgg
     3301 tcgggcgctg cgggtcgtgg ggcgggcgtc aggcaccggg cttgcgggtc atgcaccagg
     3361 tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga
     3421 aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggccg
     3481 cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga
     3541 cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt
     3601 ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct
     3661 cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc
     3721 cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt
     3781 gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt
     3841 agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg cgggtggcga
     3901 ggcgcaccgt gggcttgtac tcggtcatgg taagctccgg tggatccccc tggggagaga
     3961 ggtcggtgat tcggtcaacg agggagccga ctgccgacgt gcgctccgga ggcttgcaga
     4021 atgcggaaca ccgcgcgggc aggaacaggg cccacactac cgccccacac cccgcctccc
     4081 gcaccgcccc ttcccggccg ctgctctcgg cgcgccctgc tgagcagccg ctattggcca
     4141 cagcccatcg cggtcggcgc gctgccattg ctccctggcg ctgtccgtct gcgagggtac
     4201 tagtgagacg tgcggcttcc gtttgtcacg tccggcacgc cgcgaaccgc aaggaacctt
     4261 cccgacttag gggcggagca ggaagcgtcg ccggggggcc cacaagggta gcggcgaaga
     4321 tccgggtgac gctgcgaacg gacgtgaaga atgtgcgaga cccagggtcg gcgccgctgc
     4381 gtttcccgga accacgccca gagcagccgc gtccctgcgc aaacccaggg ctgccttgga
     4441 aaaggcgcaa ccccaacccc ctcgagccgc ggccaaagtg gatctctgct gtccctgtaa
     4501 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     4561 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     4621 ttaaaattgt ggatgaatac tgccatttgt ctcgaggtcg agaattaaaa acctcgaaga
     4681 catcggcaga aactcgagtt tctgccgatg tcttcgaggc cggtgtttcg tcctttccac
     4741 aagatatata aagccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc
     4801 attttaaaac ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac
     4861 gtattttgta ctaatatctt tgtgtttaca gtcaaattaa ttccaattat ctctctaaca
     4921 gccttgtatc gtatatgcaa atatgaagga atcatgggaa ataggccctc ggtgaagggg
     4981 gcggccgctc gaggctagtc tcgtgatcga taccgtcgag atccgttcac taatcgaatg
     5041 gatctgtctc tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc
     5101 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     5161 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     5221 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     5281 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     5341 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     5401 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     5461 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     5521 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     5581 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     5641 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     5701 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     5761 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     5821 ttatataatt cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc
     5881 ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct tgtctaaagc
     5941 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
     6001 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
     6061 ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc gttctagctc
     6121 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
     6181 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
     6241 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
     6301 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt
     6361 ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa
     6421 aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga
     6481 agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc
     6541 tcccaggctc agatctggtc taaccagaga gacccgttta tgtatcgagc taggcactta
     6601 aatacaatat ctctgcaatg cggcaattca gtggttcgtc caatccatgt cagacccgtc
     6661 tgttgccttc ctaataaggc acgatcgtac caccttactt ccaccaatcg gcatgcacgg
     6721 tgctttttct ctccttgtaa ggcatgttgc taactcatcg ttaccatgtt gcaagactac
     6781 aagagtattg cataagacta cattaagctt gcagctccag cttttgttcc ctttagtgag
     6841 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6901 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6961 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7021 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7081 ttg
//
