LOCUS       sequence-8749-ex        8245 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 8245)
  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 8245)
  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-PGK-H2Kb2
FEATURES             Location/Qualifiers
     source          1..8245
                     /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(4450..4540)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(4510..4529)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(5204..5220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5204..5220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5217..5239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5228..5244
                     /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(5252..5258,5259..5276,5277..5282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5297..5318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5435..5452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5606..6194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5686..5705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6365..7156,7157..7225))
                     /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     6988..7007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7226..7330)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7398..7416
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7454..7476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7576..7595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7789..7811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7803..7820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7804..7820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8023..8042
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8244..8245,1..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 ttcgtttaaa
     4441 cgggctcgag tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt
     4501 ttatggtgat ttcccagaac acatagcgac atgcaaatat tgggatccgc tagcttaagt
     4561 aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg
     4621 ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct
     4681 gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc
     4741 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta
     4801 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     4861 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     4921 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     4981 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta
     5041 acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc
     5101 tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta
     5161 tggacagcgc agaagagctg gggagaatta attcgtaatc atggtcatag ctgtttcctg
     5221 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7201 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7261 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7321 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7381 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7441 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7501 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7561 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7621 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     7681 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     7741 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     7801 gttgtaaaac gacggccagt gccaagctcg ctctccctta tgcgactcct gcattaggaa
     7861 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     7921 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     7981 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     8041 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     8101 gatcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact
     8161 caacaatatc accagctgaa gcctatagag tacgagccat agataaaata aaagatttta
     8221 tttagtctcc agaaaaaggg gggaa
//
