LOCUS       sequence-224269-        7903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of mouse LIN28A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7903)
  AUTHORS   Baer C, Squadrito ML, Laoui D, Thompson D, Hansen SK, Kiialainen A, 
            Hoves S, Ries CH, Ooi CH, De Palma M
  TITLE     Suppression of microRNA activity amplifies IFN-gamma-induced 
            macrophage activation and promotes anti-tumour immunity.
  JOURNAL   Nat Cell Biol. 2016 Jul;18(7):790-802. doi: 10.1038/ncb3371. Epub 
            2016 Jun 13.
  PUBMED    27295554
REFERENCE   2  (bases 1 to 7903)
  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: "10.1038/ncb3371"; 
            journalName: "Nat Cell Biol"; date: "2016-07"; volume: "18"; issue: 
            "7"; pages: "790-802"
COMMENT     Sequence Label: LV-LIN28A
FEATURES             Location/Qualifiers
     source          1..7903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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     complement(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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)."
     primer_bind     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        2292..2671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2672..2875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2826..2846
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2890..3070
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3117..3242
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3735..3968
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4153..4197
                     /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    4464..4581
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4627..5137
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5056..5074
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     primer_bind     complement(6062..6082)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6480..6491)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6640..6873
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6945..7066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6982..7001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7036..7055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7106..7241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7261..7280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7262..7280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7290..7307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7290..7306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7299..7321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7448..7876
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7535..7554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7745..7766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttggcc
     2221 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
     2281 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2341 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2401 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2461 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2521 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2581 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2641 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2701 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2761 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2821 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     2881 agtgaaccgg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     2941 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     3001 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     3061 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     3121 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     3181 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     3241 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     3301 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     3361 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     3421 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     3481 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     3541 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     3601 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     3661 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     3721 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     3781 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     3841 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     3901 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     3961 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     4021 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     4081 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     4141 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     4201 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     4261 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     4321 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     4381 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     4441 ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca
     4501 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     4561 attacaaaaa ttcaaaattt tatcgatcac gagactagcc tcgagaagct tgatatcgaa
     4621 ttccacgggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
     4681 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
     4741 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
     4801 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
     4861 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
     4921 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
     4981 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
     5041 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
     5101 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccctttg cctccggact
     5161 tctctggggc cagcagccgc ccgacctggg gcccggggcc acgggctcag cagacgacca
     5221 tgggctcggt gtccaaccag cagtttgcag gtggctgcgc caaggcagcg gagaaggcgc
     5281 cagaggaggc gccgcctgac gcggcccgag cggcagacga gccgcagctg ctgcacgggg
     5341 ccggcatctg taagtggttc aacgtgcgca tggggttcgg cttcctgtct atgaccgccc
     5401 gcgccggggt cgcgctcgac cccccggtgg acgtctttgt gcaccagagc aagctgcaca
     5461 tggaagggtt ccgaagcctc aaggagggtg aggcggtgga gttcaccttt aagaagtctg
     5521 ccaagggtct ggaatccatc cgtgtcactg gccctggtgg tgtgttctgt attggaagtg
     5581 agcggcggcc aaaggggaag aacatgcaga agcgaagatc caaaggagac aggtgctaca
     5641 actgcggtgg gctagaccat catgccaagg aatgcaagct gccaccccag cccaagaagt
     5701 gccacttttg ccaaagcatc aaccatatgg tggcctcgtg tccactgaag gcccagcagg
     5761 gccccagttc tcagggaaag cctgcctact tccgggagga agaggaagag atccacagcc
     5821 ctgccctgct cccagaagcc cagaattgag gcccaggagt cagggttatt ctttggctaa
     5881 tggggagttt aaggaaagag gcatcaatct gcagagtgga gaaagtgggg gtaagggtgg
     5941 gttgcgtggg tagcttgcac tgccgtgtct caggccgggg ttcccagtgt caccctgtct
     6001 ttgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     6061 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     6121 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     6181 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     6241 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     6301 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6361 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     6421 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6481 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctcta gagcctcttc
     6541 cgcgtcttcg ccttcccggg tcgagctcgg tacctttaag accaatgact tacaaggcag
     6601 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6661 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6721 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6781 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6841 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6901 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     7021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     7081 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     7141 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     7201 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     7261 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     7321 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     7381 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     7441 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7501 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7561 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7621 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7681 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7741 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7801 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     7861 aaatttaacg cgaattttaa caaaatatta acgcttacaa ttt
//
