LOCUS       sequence-8751-ex        8309 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 8309)
  AUTHORS   Li QJ, Chau J, Ebert PJ, Sylvester G, Min H, Liu G, Braich R, 
            Manoharan M, Soutschek J, Skare P, Klein LO, Davis MM, Chen CZ
  TITLE     miR-181a is an intrinsic modulator of T cell sensitivity and 
            selection.
  JOURNAL   Cell. 2007 Apr 6. 129(1):147-61.
  PUBMED    17382377
REFERENCE   2  (bases 1 to 8309)
  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: "Cell. 2007 
            Apr 6. 129(1):147-61."
COMMENT     Sequence Label: MDH1-shSHP2-3-H2Kb2
FEATURES             Location/Qualifiers
     source          1..8309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1716..1732
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     promoter        1796..2296
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     2229..2248
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     misc_feature    3414..3875
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3470..3487)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3697..3716
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     promoter        complement(4514..4604)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(4574..4593)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(5268..5284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5281..5303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5292..5308
                     /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(5316..5322,5323..5340,5341..5346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5499..5516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6429..7220,7221..7289))
                     /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     7052..7071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7462..7480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7518..7540)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7640..7659
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7853..7875
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7867..7884
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7868..7884
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8087..8106
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8308..8309,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatcttata tggggcaccc ccgccccttg
     1441 taaacttccc tgaccctgac atgacaagag ttactaacag cccctctctc caagctcact
     1501 tacaggctct ctacttagtc cagcacgaag tctggagacc tctggcggca gcctaccaag
     1561 aacaactgga ccgaccggtg gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc
     1621 gccgacacca gactaagaac ctagaacctc gctggaaagg accttacaca gtcctgctga
     1681 ccacccccac cgccctcaaa gtagacggca tcgcagcttg gatacacgcc gcccacgtga
     1741 aggctgccga ccccgggggt ggaccatcct ctagactgcc ggatcaattc ctaccgggta
     1801 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcggcccc gctgggcact
     1861 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     1921 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     1981 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     2041 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     2101 agcggccaat agcagctttg gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     2161 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     2221 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     2281 atctccgggc ctttcgacct gcaccatggt accgtgcacg ctgctcctgc tgttggcggc
     2341 cgccctggct ccgactcaga cccgcgcggg cccacactcg ctgaggtatt tcgtcaccgc
     2401 cgtgtcccgg cccggcctcg gggagccccg gtacatggaa gtcggctacg tggacgacac
     2461 ggagttcgtg cgcttcgaca gcgacgcgga gaatccgaga tatgagccgc gggcgcggtg
     2521 gatggagcag gaggggcccg agtattggga gcgggagaca cagaaagcca agggcaatga
     2581 gcagagtttc cgagtggacc tgaggaccct gctcggctac tacaaccaga gcaagggcgg
     2641 ctctcacact attcaggtga tctctggctg tgaagtgggg tccgacgggc gactcctccg
     2701 cgggtaccag cagtacgcct acgacggctg cgattacatc gccctgaacg aagacctgaa
     2761 aacgtggacg gcggcggaca tggcggcgct gatcaccaaa cacaagtggg agcaggctgg
     2821 tgaagcagag agactcaggg cctacctgga gggcacgtgc gtggagtggc tccgcagata
     2881 cctgaagaac gggaacgcga cgctgctgcg cacagattcc ccaaaggccc atgtgaccca
     2941 tcacagcaga cctgaagata aagtcaccct gaggtgctgg gccctgggct tctaccctgc
     3001 tgacatcacc ctgacctggc agttgaatgg ggaggagctg atccaggaca tggagcttgt
     3061 ggagaccagg cctgcagggg atggaacctt ccagaagtgg gcatctgtgg tggtgcctct
     3121 tgggaaggag cagtattaca catgccatgt gtaccatcag gggctgcctg agcccctcac
     3181 cctgagatgg gagcctcctc catccactgt ctccaacatg gcgaccgttg ctgttctggt
     3241 tgtccttgga gctgcaatag tcactggagc tgtggtggct tttgtgatga agatgagaag
     3301 gagaaacaca ggtggaaaag gagggtagta acgcgtgaga tacgtaaaac aattgaagat
     3361 ctagtcgacc aattccggtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3421 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3481 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     3541 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     3601 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     3661 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     3721 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     3781 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg
     3841 gggacgtggt tttcctttga aaaacacgat gataatacca tgaaaacatt taacatttct
     3901 caacaggatc tagaattagt agaagtagcg acagagaaga ttacaatgct ttatgaggat
     3961 aataaacatc atgtgggagc ggcaattcgt acgaaaacag gagaaatcat ttcggcagta
     4021 catattgaag cgtatatagg acgagtaact gtttgtgcag aagccattgc gattggtagt
     4081 gcagtttcga atggacaaaa ggattttgac acgattgtag ctgttagaca cccttattct
     4141 gacgaagtag atagaagtat tcgagtggta agtccttgtg gtatgtgtag ggagttgatt
     4201 tcagactatg caccagattg ttttgtgtta atagaaatga atggcaagtt agtcaaaact
     4261 acgattgaag aactcattcc actcaaatat acccgaaatt aatgagttaa cttaattaag
     4321 aattgcggcc gcgtcgacct gcagccaagc ttatcgataa aataaaagat tttatttagt
     4381 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagaa ttcaaaaaac
     4441 tacatgatat gcgtgggcaa gaagtcatct cttgcgtgac ttcttgccca cgcatatcat
     4501 gtagtgggct cgagtggtct catacagaac ttataagatt cccaaatcca aagacatttc
     4561 acgtttatgg tgatttccca gaacacatag cgacatgcaa atattgggat ccgctagctt
     4621 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     4681 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     4741 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     4801 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaaat gaccctgtgc
     4861 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4921 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     4981 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     5041 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     5101 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     5161 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     5221 attatggaca gcgcagaaga gctggggaga attaattcgt aatcatggtc atagctgttt
     5281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     7441 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     7501 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     7561 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     7621 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     7681 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     7741 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7801 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     7861 cgacgttgta aaacgacggc cagtgccaag ctcgctctcc cttatgcgac tcctgcatta
     7921 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     7981 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     8041 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     8101 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     8161 agaggatcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     8221 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     8281 tttatttagt ctccagaaaa aggggggaa
//
