LOCUS       sequence-261274-        8363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FTH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8363)
  AUTHORS   Galligan JT, Martinez-Noel G, Arndt V, Hayes S, Chittenden TW, 
            Harper JW, Howley PM
  TITLE     Proteomic analysis and identification of cellular interactors of the
            giant ubiquitin ligase HERC2.
  JOURNAL   J Proteome Res. 2015 Feb 6;14(2):953-66. doi: 10.1021/pr501005v. 
            Epub 2014 Dec 15.
  PUBMED    25476789
REFERENCE   2  (bases 1 to 8363)
  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: "10.1021/pr501005v";
            journalName: "J Proteome Res"; date: "2015-02-6- 6"; volume: "14"; 
            issue: "2"; pages: "953-66"
COMMENT     Sequence Label: p7126 pHAGE-P-CMVt-N-HA-GAW-FTH1
FEATURES             Location/Qualifiers
     source          1..8363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     752..769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1009..1030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1045..1050,1051..1068,1069..1075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1083..1099
                     /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     1088..1110
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1107..1123
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1107..1123
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1173..1192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1384..2017
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2064..2189
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2686..2919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3104..3148
                     /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    3441..3558
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3610..3913
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3914..4117
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4068..4088
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4119..4137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4140..4158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4168..4192
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4279..4288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4291..4317
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    4333..4357
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4907..4931)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4950..5449
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4985..5008)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5382..5401
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5470..6069
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5470..5489)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5966..5986
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6136..6724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6189..6209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6607..6618)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6799..7032
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7151..7169)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7237..7341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7342..7410,7411..8202)
                     /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(7560..7579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      781 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcaagct
      841 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
      901 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgtg
      961 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     1021 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     1081 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaatttcac
     1141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1201 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac
     1261 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc
     1321 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     1381 agttggaagg gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc
     1441 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     1501 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     1561 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     1621 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     1681 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     1741 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     1801 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     1861 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     1921 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     1981 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     2041 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     2101 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     2161 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     2221 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     2281 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     2341 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     2401 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     2461 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     2521 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     2581 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     2641 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     2701 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     2761 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     2821 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     2881 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     2941 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     3001 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     3061 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     3121 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     3181 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     3241 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     3301 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     3361 agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc gaattcacaa
     3421 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     3481 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     3541 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggactagt
     3601 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3661 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3721 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3781 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3841 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3901 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     3961 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     4021 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     4081 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct
     4141 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag
     4201 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
     4261 tctagcgttt aaacgctagc caccatggcg tacccatacg atgttccaga ttacgctgct
     4321 cgaggttatc aaacaagttt gtacaaaaaa gttggcatga cgaccgcgtc cacctcgcag
     4381 gtgcgccaga actaccacca ggactcagag gccgccatca accgccagat caacctggag
     4441 ctctacgcct cctacgttta cctgtccatg tcttactact ttgaccgcga tgatgtggct
     4501 ttgaagaact ttgccaaata ctttcttcac caatctcatg aggagaggga acatgctgag
     4561 aaactgatga agctgcagaa ccaacgaggt ggccgaatct tccttcagga tatcaaggaa
     4621 ccagactgtg atgactggga gagcgggcag aatgcaatgg agtgtgcatt acatttggaa
     4681 aaaaatgtga atcagtcact actggaactg cacaaactgg ccactgacaa aaatgacccc
     4741 catttgtgtg acttcattga gacacattac ctgaatgagc aggtgaaagc catcaaagaa
     4801 ttgggtgacc acgtgaccaa cttgcgcaag atgggagcgc ccgaatctgg cttggcggaa
     4861 tatctctttg acaagcacac cctgggagac agtgataatg aaagctaccc aactttcttg
     4921 tacaaagtgg ttcgataacg aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     4981 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     5041 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     5101 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     5161 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     5221 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     5281 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     5341 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     5401 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
     5461 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5521 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5581 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5641 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5701 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5761 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5821 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5881 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5941 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     6001 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     6061 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
     6121 gcatcgatag atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6181 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6241 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6301 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6361 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6421 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6481 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     6541 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6601 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6661 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6721 ccgcctgaga tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6781 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     6841 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6901 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6961 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7021 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7081 aatgaatatc agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga
     7141 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     7201 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     7261 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7321 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7381 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     7441 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     7501 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     7561 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     7621 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7681 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7741 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7801 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7861 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7921 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7981 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8041 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     8101 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8161 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     8221 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8281 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8341 tcagaccccg tagaaaagat caa
//
