LOCUS       sequence-246087-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for mouse C/EBPa; adds Flag tag to the N-terminus
            of protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  AUTHORS   Roe JS, Mercan F, Rivera K, Pappin DJ, Vakoc CR
  TITLE     BET Bromodomain Inhibition Suppresses the Function of Hematopoietic 
            Transcription Factors in Acute Myeloid Leukemia.
  JOURNAL   Mol Cell. 2015 May 12. pii: S1097-2765(15)00267-1. doi: 
            10.1016/j.molcel.2015.04.011.
  PUBMED    25982114
REFERENCE   2  (bases 1 to 6534)
  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: "Mol Cell. 
            2015 May 12. pii: S1097-2765(15)00267-1. doi: 
            10.1016/j.molcel.2015.04.011."
COMMENT     Sequence Label: pcDNA3 Flag C/EBPa
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1796..1819
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1853..1871)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1854..1871)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1891..1908)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1897..2121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2167..2595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2254..2273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2464..2485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2604..2624)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2609..2938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2789..2924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2851..2870
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3005..3799
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3059..3078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3669..3688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3973..4094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4010..4029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4064..4083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4143..4159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4143..4159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4156..4178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4167..4183
                     /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(4191..4197,4198..4215,4216..4221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4236..4257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4374..4391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4545..5133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4625..4644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5304..6095,6096..6164))
                     /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     5927..5946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6165..6269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6344..6363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca ccatggagtc
      721 ggccgacttc tacgaggtgg agccgcggcc cccgatgagc agtcacctcc agagcccccc
      781 gcacgcgccc agcaacgccg cctttggctt tccccggggc gcgggccccg cgccgccccc
      841 agccccacct gccgccccgg agccgctggg cggcatctgc gagcacgaga cgtctataga
      901 catcagcgcc tacatcgacc cggccgcctt caacgacgag ttcctggccg acctcttcca
      961 gcacagccga cagcaggaga aggccaaggc ggcggcgggc cccgcgggtg gcggcggtga
     1021 ctttgactac ccgggagccc cggcgggccc cggcggcgcg gtcatgtccg cgggggcgca
     1081 cgggccccct cccggctacg gctgtgcggc ggccggctac ctggacggca ggctggagcc
     1141 cctgtacgag cgcgtcgggg cgcccgcgct acggccgctg gtgatcaaac aagagccccg
     1201 cgaggaggac gaggcgaagc agctggcgct ggccggcctc ttcccctacc agccaccgcc
     1261 gccaccgcca ccgccgcacc cgcacgcgtc tcccgcgcac ctggccgccc cccacttgca
     1321 gttccagatc gcgcactgcg gccagaccac catgcacctg cagcctggcc accccacacc
     1381 gccgcccacg cccgtgccca gcccgcacgc tgcgcccgcc ttgggtgctg cgggcctgcc
     1441 tggccccggg agcgcgctca agggcttggc cggtgcgcac cccgacctcc gcacgggagg
     1501 cggcggcggt ggcagcggtg ccggtgcggg caaagccaag aagtcggtgg acaagaacag
     1561 caacgagtac cgggtacggc gggaacgcaa caacatcgcg gtgcgcaaga gccgagataa
     1621 agccaaacaa cgcaacgtgg agacgcaaca gaaggtgctg gagttgacca gtgacaatga
     1681 ccgcctgcgc aagcgggtgg aacagctgag ccgtgaactg gacacgctgc ggggcatctt
     1741 ccgccagctg cctgagagct ccttggtcaa ggccatgggc aactgcgcgg gaggcgacta
     1801 caaggacgac gatgacaagt gatgactcga gcatgcatct agagggccct attctatagt
     1861 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     1921 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1981 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2041 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2101 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2161 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2221 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2281 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2341 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2401 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2461 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2521 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2581 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2641 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2701 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2761 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2821 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     2881 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2941 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3001 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3061 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3181 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3301 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3781 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3961 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4021 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4081 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4141 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4201 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4261 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4321 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4981 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5281 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5341 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5401 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5461 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5521 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5581 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5641 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5701 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5761 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5821 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5881 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5941 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6001 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6061 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6121 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6301 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6361 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6421 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     6481 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
