LOCUS       sequence-429067-        8164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration of CEBPa AroLITE mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8164)
  AUTHORS   Naderi J, Magalhaes AP, Kibar G, Stik G, Zhang Y, Mackowiak SD, 
            Wieler HM, Rossi F, Buschow R, Christou-Kent M, Alcoverro-Bertran M,
            Graf T, Vingron M, Hnisz D
  TITLE     An activity-specificity trade-off encoded in human transcription 
            factors.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1309-1321. doi: 
            10.1038/s41556-024-01411-0. Epub 2024 Jul 5.
  PUBMED    38969762
REFERENCE   2  (bases 1 to 8164)
  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/s41556-024-01411-0"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1309-1321"
COMMENT     Sequence Label: pHAGE-mEGFP-hCEBPa_AroLITE
FEATURES             Location/Qualifiers
     source          1..8164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2251..2269,2270..2292,2293..2311,2312..2334,2335..2353,
                     2354..2376,2377..2395,2396..2418,2419..2437,2438..2460,
                     2461..2479,2480..2502,2503..2521)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     2589..2613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3358..3369
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             join(3876..3878,3879..4592)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3918..3939)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4179..4198)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4526..4547
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4606..5194
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4659..4679)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5077..5088)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5342..5522
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5569..5587)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5655..5759
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5760..5828,5829..6620)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5978..5997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6791..7379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7533..7550
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7790..7811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7826..7831,7832..7849,7850..7856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7864..7880
                     /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     7869..7891
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7888..7904
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7888..7904
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7954..7973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca
     2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2461 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc atgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc caccatgcgc gggcgcgggc
     2701 gagcagggtc tccgggtggg cggcggcgcc ggccagccca agcaggcggg agaaggggta
     2761 gcccctgtcg agaaaattcc aatagtccaa tggaaagtgc agatgccgcc gaagccgaac
     2821 ccaggccacc aatgtcatca catctccaat ccccacccca tgctccatca agtgcagcag
     2881 ccggtgcccc aaggggcgct gggcctgctc aaccgccagc acccccagct gctcccgaac
     2941 cactcggcgg aatttgtgaa catgaaacca gcattgatat atctgctgca attgatccag
     3001 cagcggcaaa tgatgaagcc ctcgcagatc ttgcacaaca tagtaggcaa caagagaaag
     3061 ctaaagctgc ggtcggacct accggcggag gaggaggagg tgatgcggat gcacccggtg
     3121 cacctgcagg gcctggtggg gcagttatgc caggcggggc ccatggacct cctccgggag
     3181 caggatgtgc tgcggctggg gcactcgatg gaagattgga accattggca gaacgagtag
     3241 gagcaccagc ccttagacct cttgtaatta aacaagaacc tagggaagaa gacgaggcga
     3301 aacaactcgc acttgctggt ctggcccccg cccaacctcc tccaccccca ccacctagtc
     3361 atcctcatcc tcatccccca ccagctcatc ttgcagctcc tcatctccaa gcgcaaatag
     3421 ctcattgtgg acaaactact atgcatcttc aacctggcca tccgacaccc ccaccaaccc
     3481 cggttccaag tccacatcct gcacctgctc tgggagcagc tggactccct ggtccggggt
     3541 ctgccttgaa aggcctcggt gcagcacatc cagatctgag agcttcaggc gggtccgggg
     3601 caggaaaagc aaagaaaagt gtcgataaga atagtaatga atatagagtt cgccgtgaaa
     3661 ggaataatat agccgtccgg aaatctaggg ataaagctaa acaacgaaat gtcgaaaccc
     3721 aacaaaaggt tctcgaactc actagcgata acgataggct cagaaaacgt gtagagcaac
     3781 tctctcggga gttggatacc ttgagaggca tcttccgcca gctgccagag agctccttgg
     3841 tcaaggccat gggcaactgc gcgggctcga gcggcgtgag caagggcgag gagctgttca
     3901 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3961 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4021 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4081 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4141 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     4201 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     4261 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     4321 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     4381 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4441 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     4501 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4561 tcactctcgg catggacgag ctgtacaagt aaatcgatag atcctaatca acctctggat
     4621 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4681 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4741 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     4801 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4861 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4921 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4981 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5041 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5101 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5161 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     5221 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5281 tcactcccaa cgaagacaag atcacctgca ggacaggcgc gccctgcttt ttgcttgtac
     5341 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5401 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5461 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     5521 cacccgggcg attaaggaaa gggctagatc attcttgaag acgaaagggc ctcgtgatac
     5581 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5761 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5821 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5881 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5941 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6001 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6061 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6301 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6361 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6601 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6661 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6721 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6901 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7261 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7321 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7381 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7441 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7501 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7561 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaagc
     7621 tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7681 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccgt
     7741 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7801 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     7861 gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttca
     7921 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7981 cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca
     8041 ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt
     8101 cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag
     8161 tagt
//
