LOCUS       sequence-358572-        3279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Erg enhancer in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3279)
  AUTHORS   Wang D, Tanaka-Yano M, Meader E, Kinney MA, Morris V, Lummertz da 
            Rocha E, Liu N, Liu T, Zhu Q, Orkin SH, North TE, Daley GQ, Rowe RG
  TITLE     Developmental maturation of the hematopoietic system controlled by a
            Lin28b-let-7-Cbx2 axis.
  JOURNAL   Cell Rep. 2022 Apr 5;39(1):110587. doi: 
            10.1016/j.celrep.2022.110587.
  PUBMED    35385744
REFERENCE   2  (bases 1 to 3279)
  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 Rep.";
            date: "2022-04-5"; volume: "39(1):110587. doi"; pages: " 
            10.1016/j.celrep.2022.110587"
COMMENT     Sequence Label: pNL3.1-Erg
FEATURES             Location/Qualifiers
     source          1..3279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        205..236
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      263..272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             269..784
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    833..954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(844..863)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     898..917
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1202..1219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1373..1961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1453..1472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2161..2952,2953..3021))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3126..3174
                     /note="synthetic polyadenylation signal"
     misc_feature    3188..3279
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3228..3247
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta ggagtggacc acagtgcctg ccccaatcca
       61 atcgcgcgga acttgagact gcggtacata gggaagcagg ctacccgggc ctgatctggg
      121 gcacaattcc cttccgctcg actcgtgact cgagccccgg gctaccttgc cagcagagtc
      181 ttgcccaaca cccagcttag acactagagg gtatataatg gaagctcgac ttccagcttg
      241 gcaatccggt actgttggta aagccaccat ggtcttcaca ctcgaagatt tcgttgggga
      301 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag
      361 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga
      421 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca
      481 aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa
      541 ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta
      601 tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg
      661 gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct
      721 gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc
      781 gtaaggccgc gactctagag tcggggcggc cggccgcttc gagcagacat gataagatac
      841 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
      901 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
      961 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1021 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     1081 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     1141 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     1201 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1261 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1321 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1381 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1441 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1501 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1561 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1621 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1681 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1741 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1801 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1861 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1921 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1981 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2041 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2101 aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg
     2161 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
     2221 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
     2281 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
     2341 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
     2401 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
     2461 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
     2521 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
     2581 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
     2641 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
     2701 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
     2761 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
     2821 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
     2881 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
     2941 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
     3001 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
     3061 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
     3121 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     3181 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     3241 ctgtccccag tgcaagtgca ggtgccagaa catttctct
//
