LOCUS       sequence-382004-        6336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ZNF354C with mutated KRAB domain (E25A, W26A mutation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  AUTHORS   Oo JA, Irmer B, Gunther S, Warwick T, Palfi K, Izquierdo Ponce J, 
            Teichmann T, Pfluger-Muller B, Gilsbach R, Brandes RP, Leisegang MS
  TITLE     ZNF354C is a transcriptional repressor that inhibits endothelial 
            angiogenic sprouting.
  JOURNAL   Sci Rep. 2020 Nov 5;10(1):19079. doi: 10.1038/s41598-020-76193-0.
  PUBMED    33154469
REFERENCE   2  (bases 1 to 6336)
  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: "Sci Rep."; 
            date: "2020-11-5"; volume: "10(1):19079. doi"; pages: " 
            10.1038/s41598-020-76193-0"
COMMENT     Sequence Label: pCMV6-ZNF354C_mutant_E25A-W26A
FEATURES             Location/Qualifiers
     source          1..6336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2502..2531
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2550..2573
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2593..2609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2593..2609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2620..3242
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2819..2838)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3391..3979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3471..3490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4308..4355
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4380..4399
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4587..5381)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4587..5381)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4698..4717)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5308..5327
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5416..5773)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5430..5565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5582..5773
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5730..5750
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5775..5879)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5906..6334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5993..6012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6203..6224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggctgtggat ctgctgtctg ctcaggagcc tgtgacattc agggatgtgg
     1081 ccgtgttctt cagccaggac gcggcgttgc acctggactc tgcccagagg gccttgtacc
     1141 gggaggtgat gctggagaac tacagcagcc tggtctcact ggggattcca ttttcaatgc
     1201 caaagttgat tcatcagttg cagcaaggag aagatccctg catggtggaa agagaagtcc
     1261 cttcagatac ccgtctaggt ttcaagactt ggcttgaaac agaagcattg cctcatagac
     1321 aggacatttt tatagaagaa acatctcagg gaatggtaaa gaaagaatcc attaaggatg
     1381 gtcactggga cattaacttt gaagaagctg tggaatttga gagcgagata gaagaagagc
     1441 aagagaagaa acctcttaga caaatgatag attcgcatga gaaaaccatc agtgaagatg
     1501 gaaaccatac aagtcttgaa ttggggaaaa gcttatttac aaatacagct cttgtcacac
     1561 aacagagtgt tcctatagaa aggataccca atatgtatta tacatttggg aaagatttta
     1621 aacagaattt tgatctcatg aaatgcttcc agatttaccc aggaggaaaa cctcacatct
     1681 gtaatgaatg tgggaagagc ttcaagcaga atctgcatct tattgaacat cagagaattc
     1741 atacaggtga gaaaccctac aaatgtaatg agtgtaaaaa aaccttcagc cacagatcat
     1801 cccttctttc tcatcagaga attcatactg gagagaaacc ttacaagtgt aatgaatgtg
     1861 agaaggcatt tagcaacagt tcaaccctta tcaaacatct gagagtgcat actggagaga
     1921 aaccgtatcg atgtagggaa tgtggtaaag cctttagcca gtgttcaacc ctcactgtac
     1981 atcagagaat tcatactgga gagaaactct ataaatgcgg cgaatgtgag aaggccttca
     2041 actgtagagc aaaacttcac aggcatcaaa gaatccatac aggtgagaaa ccctataaat
     2101 gtagtgagtg tgggaaggga tacagccagt ttacatctct agctgaacat cagaggtttc
     2161 atactggaga acaactgtat acatgcttgg aatgtgggag aaccttcaca cgtattgtaa
     2221 cccttatcga acatcagcga attcacactg gacaaaaacc ttatcagtgc aacgaatgtg
     2281 agaaagcctt caaccagtat tcatccttta atgaacatcg gaaaattcat actggggaaa
     2341 aactttatac atgtgaggaa tgtgggaaag cctttggttg caaatctaac ctttataggc
     2401 atcagagaat tcatactgga gagaaaccgt atcagtgtaa tcagtggaaa ggccttcagc
     2461 cagtattcat ttttaaccga acaacgcgta cgcggccgct cgagcagaaa ctcatctcag
     2521 aagaggatct ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa
     2581 cggccggccg cggtcatagc tgtttcctga acagatcccg ggtggcatcc ctgtgacccc
     2641 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     2701 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     2761 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     2821 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     2881 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     2941 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3001 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3061 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     3121 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     3181 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     3241 ttgggtacgc ggccagcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3301 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3361 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3421 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3481 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3541 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3601 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3661 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3721 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3781 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3841 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3901 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3961 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4021 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4081 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     4141 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     4201 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     4261 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     4321 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     4381 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     4441 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     4501 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     4561 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     4621 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4681 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcgatccgcc
     4741 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4801 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     4861 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     4921 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     4981 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     5041 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     5101 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     5161 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     5221 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     5281 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     5341 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     5401 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     5461 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     5521 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     5581 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5641 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     5701 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     5761 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5821 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5881 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6301 aatgagctga tttaacaaaa atttaacgcg aatttt
//
