LOCUS       sequence-8750-ex        8312 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 8312)
  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 8312)
  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-shDusp5-2-H2Kb2
FEATURES             Location/Qualifiers
     source          1..8312
                     /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(4517..4607)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(4577..4596)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(5271..5287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5271..5287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5284..5306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5295..5311
                     /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(5319..5325,5326..5343,5344..5349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5364..5385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5502..5519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5673..6261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5753..5772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6432..7223,7224..7292))
                     /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     7055..7074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7293..7397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7465..7483
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7521..7543)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7643..7662
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7856..7878
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7870..7887
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7871..7887
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8090..8109
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8311..8312,1..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 ttcaaaaatt
     4441 ataaacgtgg agctggctgc ctccccttgt ttcagaatcc aaggggaggc agccagctcc
     4501 acgtttatgg gctcgagtgg tctcatacag aacttataag attcccaaat ccaaagacat
     4561 ttcacgttta tggtgatttc ccagaacaca tagcgacatg caaatattgg gatccgctag
     4621 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     4681 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     4741 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     4801 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa aatgaccctg
     4861 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     4921 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg
     4981 cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     5041 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     5101 atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa
     5161 gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag
     5221 ttcattatgg acagcgcaga agagctgggg agaattaatt cgtaatcatg gtcatagctg
     5281 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5341 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5401 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5461 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7261 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7321 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7381 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     7441 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     7501 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     7561 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     7621 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     7681 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     7741 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7801 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7861 tcacgacgtt gtaaaacgac ggccagtgcc aagctcgctc tcccttatgc gactcctgca
     7921 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     7981 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     8041 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     8101 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     8161 cgtagaggat cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     8221 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     8281 gattttattt agtctccaga aaaagggggg aa
//
