LOCUS       sequence-333887-        8687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses AMOT(A112K) 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 Dec 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(A112K)
FEATURES             Location/Qualifiers
     source          1..8687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        120..449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      300..435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     362..381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             516..1310
                     /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(570..589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1180..1199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1484..1605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1521..1540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1575..1594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1654..1670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1654..1670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1667..1689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1678..1694
                     /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(1702..1708,1709..1726,1727..1732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1747..1768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1885..1902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2056..2644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2136..2155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2815..3606,3607..3675))
                     /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     3438..3457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3676..3780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3855..3874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4046..4425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4426..4629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4580..4600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4674..4693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4674..4692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4715..4741
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4745..4771
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5930..5950
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             7480..7497
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(8051..8069)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8052..8069)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    8095..8319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8365..8687,1..106)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8452..8471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8662..8683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
       61 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
      121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
      421 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
      481 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
      541 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      601 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
      661 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
      721 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      781 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      841 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      901 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      961 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1021 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     1081 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1141 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1201 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1261 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     1321 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     1381 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1441 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     1501 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1561 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1621 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1681 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2461 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3661 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3781 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     3841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     3901 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     3961 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     4021 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     4081 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     4141 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     4201 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     4261 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     4321 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     4381 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     4441 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4501 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     4561 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4621 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
     4681 actcactata gggagaccca agcttgccat ggcctaccca tacgatgttc ctgactatgc
     4741 gggctatccc tatgacgtcc cggactatgc aggatccatg agaaattctg acgaacagcc
     4801 aagtggaggg accacggtat tgcagcgttt gctacaagag cagcttcgct atggcaatcc
     4861 tagtgagaat cgcagcctgc ttgccataca ccagcaagcc acagggaatg gccctccttt
     4921 ccccagtggc agtgggaacc cgggccctca gagtgatgtg ttgagtcccc aagaccacca
     4981 ccaacagctt gtggctcatg ctgctcgaca agaaccccag gggcaggaaa tccagtcaga
     5041 aaacctcatc atggagaagc agctgtctcc tcgaatgcaa aataatgaag aactcccgac
     5101 ctatgaagaa aagaaggtcc agtcccagta ctttcggggc caacagcatg ccagtgttgg
     5161 agctgccttc tatgtcactg gagtcaccaa ccagaagatg aggactgagg gacgcccatc
     5221 agttcagcgg ctcaatcctg gaaagatgca ccaagatgag ggactcagag accttaagca
     5281 agggcatgtc cgttccttga gtgaacgact aatgcagatg tcactggcca ccagtggagt
     5341 taaggcccat ccacctgtta ccagtgctcc cctctcccca ccacaaccca atgacctcta
     5401 caagaatccc acaagttcca gtgaattcta caaggcccaa gggccacttc ctaaccagca
     5461 tagcctgaag ggcatggaac accgaggccc cccaccagaa tatcccttca agggcatgcc
     5521 accccaatct gtagtgtgca agccccaaga gccagggcac ttctatagtg agcatcgcct
     5581 gaaccagcca gggagaacag aggggcaact gatgaggtat cagcatcccc ctgagtatgg
     5641 agcagccagg ccagcgcagg acatctcatt gccattgtca gccaggaact cgcagcctca
     5701 cagccctact tcttctctga cctctggggg gtccttgccc ttgctacaat ctccaccatc
     5761 cactagattg tctcctgccc gacacccctt ggtcccaaac cagggagacc attcagctca
     5821 cctgcctagg ccgcagcagc atttccttcc taatcaggct caccaggggg atcattaccg
     5881 tctctcccaa cctggcctga gtcagcagca gcagcaacag cagcagcagc accatcatca
     5941 ccatcaccac caacaacagc agcagcagca gccacagcag cagccaggag aagcctattc
     6001 agctatgcct cgggctcagc catcctctgc ttcttatcag ccagtgccag cagacccttt
     6061 tgccattgtt tccagagccc agcagatggt tgagatcctc tcagacgaga accggaactt
     6121 gaggcaagag ttggaaggat gctatgagaa ggtggcaaga ctgcagaagg tggagacaga
     6181 aatccagcgc gtctcggagg catatgagaa cctcgtgaag tcatcctcca aaagagaggc
     6241 cctagagaaa gccatgagaa acaagctaga gggcgagatt cggaggatgc atgatttcaa
     6301 cagggatctg agagagcgtc tagagactgc caacaagcag cttgcagaga aggaatatga
     6361 ggggtcagag gacaccagaa aaaccatctc gcagctcttt gcaaaaaata aagaaagcca
     6421 gcgtgagaag gagaagctgg aagcggagct ggccactgcc cgttctacca atgaggacca
     6481 aagacgacac atcgaaatcc gagatcaggc cctgagtaat gcccaggcca aggtggtaaa
     6541 gctggaagaa gagctgaaaa agaagcaagt gtacgttgac aaggtggaga agatgcagca
     6601 ggcccttgta cagctccagg cagcatgtga aaaacgtgag cagctagagc accgtctccg
     6661 gacacgactg gagagggaac tggaatccct gagaatccag cagcgtcagg gcaactgtca
     6721 gcccaccaac gtttcagaat acaatgctgc tgcactgatg gagctccttc gggagaaaga
     6781 ggagaggatt ctggctctgg aagctgatat gacaaagtgg gagcagaaat atttggagga
     6841 gaatgtgatg agacattttg ctctggatgc tgctgcaact gtggctgctc agagggacac
     6901 aacagtcatc agtcactctc ctaacaccag ctatgacaca gctctagaag ctcgcatcca
     6961 gaaagaggag gaagaaatct tgatggccaa taagcgttgc cttgacatgg agggcaggat
     7021 taagaccctc catgcccaga ttattgagaa ggatgccatg atcaaagtac tccagcagcg
     7081 ttcccggaag gagccgagca agacagagca gctgtcgtgc atgcggccag cgaagtctct
     7141 gatgtccatt tccaatgctg gatcaggctt gctctcccac tcatccaccc tgactggctc
     7201 ccccatcatg gaagaaaagc gagacgacaa gagctggaag gggagcctag gcattctcct
     7261 gggtggagac taccgtgctg aatatgtccc ttccacaccc tcgcctgtgc caccctcgac
     7321 tcccctgctc tcggctcact ccaagacaga cagccgagac tgcagtaccc aaactgaacg
     7381 tgggacggaa tcgaacaaaa ctgcagctgt tgctcccatc tctgttcctg ctccagttgc
     7441 tgctgccgcc actgctgccg ccatcactgc cactgctgcc accatcacca ccaccatggt
     7501 agctgctgct ccagttgctg ttgctgctgc tgctgctcca gctgctgctg ctgccccgtc
     7561 tccagccact gccgctgcta ctgctgctgc tgtttctcca gctgctgctg gtcagattcc
     7621 agctgctgcc tctgttgcct cagctgctgc cgttgctcct tctgctgctg ctgctgctgc
     7681 tgttcaggtt gctccagctg ctccggctcc agttccagct ccggctctgg ttccggtccc
     7741 agctccagca gcggctcagg cttctgctcc tgctcagact caggcaccaa cttcagctcc
     7801 ggctgtggct ccaactccag ctccaactcc aactccagct gtggctcagg ctgaggttcc
     7861 tgcaagtcca gctaccggtc ctggaccaca tcgtttgtct ataccaagtt tgacctgcaa
     7921 tccagacaaa acagatgggc ctgtgttcca ctccaatact ctggaaagaa aaactcccat
     7981 tcagatcctg ggacaagagc ctgatgcaga gatggtggaa tatctcatct aactcgaggg
     8041 ggggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     8101 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     8161 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     8221 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     8281 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     8341 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     8401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     8461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     8521 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     8581 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     8641 tggagtccac gttctttaat agtggactct tgttccaaac tggaaca
//
