LOCUS       sequence-331964-        8687 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expresses AMOT(L106P) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8687)
  AUTHORS   Rheinemann L, Thompson T, Mercenne G, Paine EL, Peterson FC, Volkman
            BF, Alam SL, Alian A, Sundquist WI
  TITLE     Interactions between AMOT PPxY Motifs and NEDD4L WW Domains Function
            in HIV-1 Release.
  JOURNAL   J Biol Chem. 2021 Jul 17:100975. doi: 10.1016/j.jbc.2021.100975.
  PUBMED    34284061
REFERENCE   2  (bases 1 to 8687)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2021 Jul 17:100975. doi: 10.1016/j.jbc.2021.100975."
COMMENT     Sequence Label: pcDNA3HA-AMOT p130∆PPxY1(L106P)
FEATURES             Location/Qualifiers
     source          1..8687
                     /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             670..696
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             700..726
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1885..1905
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             3435..3452
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(4006..4024)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4007..4024)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    4050..4274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4320..4748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4617..4638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4757..4777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4762..5091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4942..5077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5004..5023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5158..5952
                     /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(5212..5231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5822..5841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6126..6247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6163..6182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6217..6236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6296..6312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6296..6312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6309..6331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6320..6336
                     /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(6344..6350,6351..6368,6369..6374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6389..6410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6527..6544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6698..7286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6778..6797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7457..8248,8249..8317))
                     /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     8080..8099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8318..8422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8497..8516)
                     /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 gccatggcct acccatacga tgttcctgac tatgcgggct atccctatga cgtcccggac
      721 tatgcaggat ccatgagaaa ttctgaagaa cagccaagtg gagggaccac ggtattgcag
      781 cgtttgctac aagagcagct tcgctatggc aatcctagtg agaatcgcag cctgcttgcc
      841 atacaccagc aagccacagg gaatggccct cctttcccca gtggcagtgg gaacccgggc
      901 cctcagagtg atgtgttgag tccccaagac caccaccaac agcttgtggc tcatgctgct
      961 cgacaagaac cccaggggca ggaaatccag tcagaaaacc tcatcatgga gaagcagctg
     1021 tctcctcgaa tgcaaaataa tgaagaaccc ccgacctatg aagaagccaa ggtccagtcc
     1081 cagtactttc ggggccaaca gcatgccagt gttggagctg ccttctatgt cactggagtc
     1141 accaaccaga agatgaggac tgagggacgc ccatcagttc agcggctcaa tcctggaaag
     1201 atgcaccaag atgagggact cagagacctt aagcaagggc atgtccgttc cttgagtgaa
     1261 cgactaatgc agatgtcact ggccaccagt ggagttaagg cccatccacc tgttaccagt
     1321 gctcccctct ccccaccaca acccaatgac ctctacaaga atcccacaag ttccagtgaa
     1381 ttctacaagg cccaagggcc acttcctaac cagcatagcc tgaagggcat ggaacaccga
     1441 ggccccccac cagaatatcc cttcaagggc atgccacccc aatctgtagt gtgcaagccc
     1501 caagagccag ggcacttcta tagtgagcat cgcctgaacc agccagggag aacagagggg
     1561 caactgatga ggtatcagca tccccctgag tatggagcag ccaggccagc gcaggacatc
     1621 tcattgccat tgtcagccag gaactcgcag cctcacagcc ctacttcttc tctgacctct
     1681 ggggggtcct tgcccttgct acaatctcca ccatccacta gattgtctcc tgcccgacac
     1741 cccttggtcc caaaccaggg agaccattca gctcacctgc ctaggccgca gcagcatttc
     1801 cttcctaatc aggctcacca gggggatcat taccgtctct cccaacctgg cctgagtcag
     1861 cagcagcagc aacagcagca gcagcaccat catcaccatc accaccaaca acagcagcag
     1921 cagcagccac agcagcagcc aggagaagcc tattcagcta tgcctcgggc tcagccatcc
     1981 tctgcttctt atcagccagt gccagcagac ccttttgcca ttgtttccag agcccagcag
     2041 atggttgaga tcctctcaga cgagaaccgg aacttgaggc aagagttgga aggatgctat
     2101 gagaaggtgg caagactgca gaaggtggag acagaaatcc agcgcgtctc ggaggcatat
     2161 gagaacctcg tgaagtcatc ctccaaaaga gaggccctag agaaagccat gagaaacaag
     2221 ctagagggcg agattcggag gatgcatgat ttcaacaggg atctgagaga gcgtctagag
     2281 actgccaaca agcagcttgc agagaaggaa tatgaggggt cagaggacac cagaaaaacc
     2341 atctcgcagc tctttgcaaa aaataaagaa agccagcgtg agaaggagaa gctggaagcg
     2401 gagctggcca ctgcccgttc taccaatgag gaccaaagac gacacatcga aatccgagat
     2461 caggccctga gtaatgccca ggccaaggtg gtaaagctgg aagaagagct gaaaaagaag
     2521 caagtgtacg ttgacaaggt ggagaagatg cagcaggccc ttgtacagct ccaggcagca
     2581 tgtgaaaaac gtgagcagct agagcaccgt ctccggacac gactggagag ggaactggaa
     2641 tccctgagaa tccagcagcg tcagggcaac tgtcagccca ccaacgtttc agaatacaat
     2701 gctgctgcac tgatggagct ccttcgggag aaagaggaga ggattctggc tctggaagct
     2761 gatatgacaa agtgggagca gaaatatttg gaggagaatg tgatgagaca ttttgctctg
     2821 gatgctgctg caactgtggc tgctcagagg gacacaacag tcatcagtca ctctcctaac
     2881 accagctatg acacagctct agaagctcgc atccagaaag aggaggaaga aatcttgatg
     2941 gccaataagc gttgccttga catggagggc aggattaaga ccctccatgc ccagattatt
     3001 gagaaggatg ccatgatcaa agtactccag cagcgttccc ggaaggagcc gagcaagaca
     3061 gagcagctgt cgtgcatgcg gccagcgaag tctctgatgt ccatttccaa tgctggatca
     3121 ggcttgctct cccactcatc caccctgact ggctccccca tcatggaaga aaagcgagac
     3181 gacaagagct ggaaggggag cctaggcatt ctcctgggtg gagactaccg tgctgaatat
     3241 gtcccttcca caccctcgcc tgtgccaccc tcgactcccc tgctctcggc tcactccaag
     3301 acagacagcc gagactgcag tacccaaact gaacgtggga cggaatcgaa caaaactgca
     3361 gctgttgctc ccatctctgt tcctgctcca gttgctgctg ccgccactgc tgccgccatc
     3421 actgccactg ctgccaccat caccaccacc atggtagctg ctgctccagt tgctgttgct
     3481 gctgctgctg ctccagctgc tgctgctgcc ccgtctccag ccactgccgc tgctactgct
     3541 gctgctgttt ctccagctgc tgctggtcag attccagctg ctgcctctgt tgcctcagct
     3601 gctgccgttg ctccttctgc tgctgctgct gctgctgttc aggttgctcc agctgctccg
     3661 gctccagttc cagctccggc tctggttccg gtcccagctc cagcagcggc tcaggcttct
     3721 gctcctgctc agactcaggc accaacttca gctccggctg tggctccaac tccagctcca
     3781 actccaactc cagctgtggc tcaggctgag gttcctgcaa gtccagctac cggtcctgga
     3841 ccacatcgtt tgtctatacc aagtttgacc tgcaatccag acaaaacaga tgggcctgtg
     3901 ttccactcca atactctgga aagaaaaact cccattcaga tcctgggaca agagcctgat
     3961 gcagagatgg tggaatatct catctaactc gagggggggc cctattctat agtgtcacct
     4021 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4081 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4141 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4201 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4261 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4741 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4861 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4921 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4981 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5041 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5101 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5161 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5221 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5281 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5341 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5401 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5461 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5521 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5581 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5641 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5701 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5761 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5821 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5881 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5941 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6001 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6061 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6121 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6241 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8461 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     8521 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8581 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     8641 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
