LOCUS       sequence-194480-        8339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8339)
  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 8339)
  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: p7127 pHAGE-P-CMVt-N-HA-GAW-FTL
FEATURES             Location/Qualifiers
     source          1..8339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     502..521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     755..772
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1012..1033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1048..1053,1054..1071,1072..1078)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1086..1102
                     /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     1091..1113
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1110..1126
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1110..1126
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1176..1195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1387..2020
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2067..2192
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2689..2922
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3107..3151
                     /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    3444..3561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3613..3916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3917..4120
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4071..4091
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4122..4140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4143..4161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4171..4195
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4282..4291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4294..4320
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    4336..4360
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4886..4910)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4929..5428
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4964..4987)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5361..5380
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5449..6048
                     /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(5449..5468)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5945..5965
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6115..6703
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6168..6188)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6586..6597)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6778..7011
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7130..7148)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7216..7320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7321..7389,7390..8181)
                     /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(7539..7558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
       61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      721 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      781 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa
      841 gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      901 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc
      961 gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     1021 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1081 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt
     1141 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1201 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     1261 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
     1321 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     1381 agtagttgga agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct
     1441 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat
     1501 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag
     1561 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg
     1621 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg
     1681 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac
     1741 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc
     1801 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag
     1861 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     1921 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     1981 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     2041 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     2101 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     2161 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     2221 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     2281 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     2341 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     2401 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     2461 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     2521 cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     2581 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     2641 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     2701 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     2761 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     2821 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     2881 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     2941 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     3001 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     3061 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     3121 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     3181 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     3241 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     3301 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     3361 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gccgaattca
     3421 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     3481 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     3541 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggact
     3601 agtggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3661 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3721 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3781 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3841 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3901 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3961 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4021 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4081 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca gtgatagaga
     4141 tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg
     4201 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
     4261 gactctagcg tttaaacgct agccaccatg gcgtacccat acgatgttcc agattacgct
     4321 gctcgaggtt atcaaacaag tttgtacaaa aaagttggca tgagctccca gattcgtcag
     4381 aattattcca ccgacgtgga ggcagccgtc aacagcctgg tcaatttgta cctgcaggcc
     4441 tcctacacct acctctctct gggcttctat ttcgaccgcg atgatgtggc tctggaaggc
     4501 gtgagccact tcttccgcga attggccgag gagaagcgcg agggctacga gcgtctcctg
     4561 aagatgcaaa accagcgtgg cggccgcgct ctcttccagg acatcaagaa gccagctgaa
     4621 gatgagtggg gtaaaacccc agacgccatg aaagctgcca tggccctgga gaaaaagctg
     4681 aaccaggccc ttttggatct tcatgccctg ggttctgccc gcacggaccc ccatctctgt
     4741 gacttcctgg agactcactt cctagatgag gaagtgaagc ttatcaagaa gatgggtgac
     4801 cacctgacca acctccacag gctgggtggc ccggaggctg ggctgggcga gtatctcttc
     4861 gaaaggctca ctctcaagca cgactaccca actttcttgt acaaagtggt tcgataacga
     4921 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     4981 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     5041 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     5101 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     5161 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     5221 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
     5281 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     5341 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     5401 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac
     5461 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     5521 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     5581 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5641 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5701 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5761 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5821 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5881 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5941 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     6001 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     6061 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgataga tcctaatcaa
     6121 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6181 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6241 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6301 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6361 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6421 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6481 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     6541 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6601 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6661 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac
     6721 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6781 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     6841 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6901 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6961 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7021 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7081 ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata
     7141 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     7201 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7261 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7321 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7381 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7441 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7501 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7561 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7621 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7681 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7741 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7801 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7861 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7921 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7981 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     8041 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8101 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     8161 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     8221 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8281 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagat
//
