LOCUS       sequence-129764-        8417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Lin28b in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8417)
  AUTHORS   Chang TC, Zeitels LR, Hwang HW, Chivukula RR, Wentzel EA, Dews M, 
            Jung J, Gao P, Dang CV, Beer MA, Thomas-Tikhonenko A, Mendell JT
  TITLE     Lin-28B transactivation is necessary for Myc-mediated let-7 
            repression and proliferation.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3384-9. doi: 
            10.1073/pnas.0808300106. Epub 2009 Feb 11.
  PUBMED    19211792
REFERENCE   2  (bases 1 to 8417)
  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.0808300106"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2009-03-3- 3"; volume: "106"; issue: "9"; pages: "3384-9"
COMMENT     Sequence Label: pMSCV-PIG-Lin28b
FEATURES             Location/Qualifiers
     source          1..8417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        2197..2696
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2232..2255)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2629..2648
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2717..3316
                     /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(2717..2736)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3213..3233
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3377..3944
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3394..3946
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3538..3555)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3765..3784
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3958..3960,3961..3963,3964..4677)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4003..4024)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4264..4283)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4611..4632
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4779..5293
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5462..5478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5462..5478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5475..5497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5486..5502
                     /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(5510..5516,5517..5534,5535..5540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5555..5576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5693..5710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5864..6452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5944..5963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6623..7414,7415..7483))
                     /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     7246..7265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7484..7588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7656..7674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7712..7734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7834..7853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8047..8069
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8197..8216
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8416..8417,1..515)
                     /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 gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga catggccgaa ggcggggcta
     1441 gcaaaggtgg tggagaagag cccgggaagc tgccggagcc ggcagaggag gaatcccagg
     1501 ttttgcgcgg aactggccac tgtaagtggt tcaatgtgcg catgggattt ggattcatct
     1561 ccatgataaa ccgagaggga agccccttgg atattccagt cgatgtattt gtacaccaaa
     1621 gcaaactatt catggaagga tttagaagcc taaaagaagg agaaccagtg gaattcacat
     1681 ttaaaaaatc ttccaaaggc cttgagtcaa tacgggtaac aggacctggt gggagcccct
     1741 gtttaggaag tgaaagaaga cccaaaggga agacactaca gaaaagaaaa ccaaagggag
     1801 atagatgcta caactgtggt ggccttgatc atcatgctaa ggaatgtagt ctacctcctc
     1861 agccaaagaa gtgccattac tgtcagagca tcatgcacat ggtggcaaac tgcccacata
     1921 aaaatgttgc acagccaccc gcgagttctc agggaagaca ggaagcagaa tcccagccat
     1981 gcacttcaac tctccctcga gaagtgggag gcgggcatgg ctgtacatca ccaccgtttc
     2041 ctcaggaggc tagggcagag atctcagaac ggtcaggcag gtcacctcaa gaagcttcct
     2101 ccacgaagtc atctatagca ccagaagagc aaagcaaaaa ggggccttca gttcaaaaaa
     2161 ggaaaaagac ataagtcgag gttaacgaat tctaccgggt aggggaggcg cttttcccaa
     2221 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     2281 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     2341 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     2401 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     2461 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     2521 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     2581 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     2641 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     2701 gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     2761 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     2821 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     2881 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     2941 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3001 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3061 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3121 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3181 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3241 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3301 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     3361 accccatgca tcgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat
     3421 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3481 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     3541 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     3601 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3661 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3721 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     3781 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     3841 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     3901 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     3961 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     4021 gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc cacctacggc
     4081 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4141 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4201 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4261 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4321 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4381 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4441 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4501 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4561 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4621 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga
     4681 attaattaag aattatcaag cttatcgata ccgtcgacct gcagccaagc ttatcgataa
     4741 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4801 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     4861 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     4921 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4981 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     5041 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     5101 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     5161 ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     5221 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     5281 gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg agggtaggag
     5341 tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac
     5401 tcctgaaata gttcattatg gacagcgcag aagagctggg gagaattaat tcgtaatcat
     5461 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5521 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6301 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     7621 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     7681 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     7741 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7801 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     7861 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     7921 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     7981 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8041 ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat
     8101 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     8161 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     8221 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt
     8281 agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata
     8341 tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct
     8401 ccagaaaaag gggggaa
//
