LOCUS       sequence-331958-        8701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses AMOT(E104K/E105K) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8701)
  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 8701)
  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(E104K/E105K)
FEATURES             Location/Qualifiers
     source          1..8701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             908..934
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             938..964
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2123..2143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             3673..3690
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(4253..4271)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4254..4271)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    4297..4521
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4567..4995
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4654..4673)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4864..4885
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5004..5024)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5009..5339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5190..5325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5252..5271
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5406..6200
                     /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(5460..5479)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6070..6089
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6374..6495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6411..6430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6465..6484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6544..6560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6544..6560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6557..6579)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6568..6584
                     /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(6592..6598,6599..6616,6617..6622))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6637..6658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6775..6792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6946..7534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7026..7045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7705..8496,8497..8565))
                     /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     8328..8347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8566..8670)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cnnagacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgc
      901 catggcctac ccatacgatg ttcctgacta tgcgggctat ccctatgacg tcccggacta
      961 tgcaggatcc atgagaaatt ctgacgaaca gccaagtgga gggaccacgg tattgcagcg
     1021 tttgctacaa gagcagcttc gctatggcaa tcctagtgag aatcgcagcc tgcttgccat
     1081 acaccagcaa gccacaggga atggccctcc tttccccagt ggcagtggga acccgggccc
     1141 tcagagtgat gtgttgagtc cccaagacca ccaccaacag cttgtggctc atgctgctcg
     1201 acaagaaccc caggggcagg aaatccagtc agaaaacctc atcatggaga agcagctgtc
     1261 tcctcgaatg caaaataata aaaaactccc gacctatgaa gaagccaagg tccagtccca
     1321 gtactttcgg ggccaacagc atgccagtgt tggagctgcc ttctatgtca ctggagtcac
     1381 caaccagaag atgaggactg agggacgccc atcagttcag cggctcaatc ctggaaagat
     1441 gcaccaagat gagggactca gagaccttaa gcaagggcat gtccgttcct tgagtgaacg
     1501 actaatgcag atgtcactgg ccaccagtgg agttaaggcc catccacctg ttaccagtgc
     1561 tcccctctcc ccaccacaac ccaatgacct ctacaagaat cccacaagtt ccagtgaatt
     1621 ctacaaggcc caagggccac ttcctaacca gcatagcctg aagggcatgg aacaccgagg
     1681 ccccccacca gaatatccct tcaagggcat gccaccccaa tctgtagtgt gcaagcccca
     1741 agagccaggg cacttctata gtgagcatcg cctgaaccag ccagggagaa cagaggggca
     1801 actgatgagg tatcagcatc cccctgagta tggagcagcc aggccagcgc aggacatctc
     1861 attgccattg tcagccagga actcgcagcc tcacagccct acttcttctc tgacctctgg
     1921 ggggtccttg cccttgctac aatctccacc atccactaga ttgtctcctg cccgacaccc
     1981 cttggtccca aaccagggag accattcagc tcacctgcct aggccgcagc agcatttcct
     2041 tcctaatcag gctcaccagg gggatcatta ccgtctctcc caacctggcc tgagtcagca
     2101 gcagcagcaa cagcagcagc agcaccatca tcaccatcac caccaacaac agcagcagca
     2161 gcagccacag cagcagccag gagaagccta ttcagctatg cctcgggctc agccatcctc
     2221 tgcttcttat cagccagtgc cagcagaccc ttttgccatt gtttccagag cccagcagat
     2281 ggttgagatc ctctcagacg agaaccggaa cttgaggcaa gagttggaag gatgctatga
     2341 gaaggtggca agactgcaga aggtggagac agaaatccag cgcgtctcgg aggcatatga
     2401 gaacctcgtg aagtcatcct ccaaaagaga ggccctagag aaagccatga gaaacaagct
     2461 agagggcgag attcggagga tgcatgattt caacagggat ctgagagagc gtctagagac
     2521 tgccaacaag cagcttgcag agaaggaata tgaggggtca gaggacacca gaaaaaccat
     2581 ctcgcagctc tttgcaaaaa ataaagaaag ccagcgtgag aaggagaagc tggaagcgga
     2641 gctggccact gcccgttcta ccaatgagga ccaaagacga cacatcgaaa tccgagatca
     2701 ggccctgagt aatgcccagg ccaaggtggt aaagctggaa gaagagctga aaaagaagca
     2761 agtgtacgtt gacaaggtgg agaagatgca gcaggccctt gtacagctcc aggcagcatg
     2821 tgaaaaacgt gagcagctag agcaccgtct ccggacacga ctggagaggg aactggaatc
     2881 cctgagaatc cagcagcgtc agggcaactg tcagcccacc aacgtttcag aatacaatgc
     2941 tgctgcactg atggagctcc ttcgggagaa agaggagagg attctggctc tggaagctga
     3001 tatgacaaag tgggagcaga aatatttgga ggagaatgtg atgagacatt ttgctctgga
     3061 tgctgctgca actgtggctg ctcagaggga cacaacagtc atcagtcact ctcctaacac
     3121 cagctatgac acagctctag aagctcgcat ccagaaagag gaggaagaaa tcttgatggc
     3181 caataagcgt tgccttgaca tggagggcag gattaagacc ctccatgccc agattattga
     3241 gaaggatgcc atgatcaaag tactccagca gcgttcccgg aaggagccga gcaagacaga
     3301 gcagctgtcg tgcatgcggc cagcgaagtc tctgatgtcc atttccaatg ctggatcagg
     3361 cttgctctcc cactcatcca ccctgactgg ctcccccatc atggaagaaa agcgagacga
     3421 caagagctgg aaggggagcc taggcattct cctgggtgga gactaccgtg ctgaatatgt
     3481 cccttccaca ccctcgcctg tgccaccctc gactcccctg ctctcggctc actccaagac
     3541 aggcagccga gactgcagta cccaaactga acgtgggacg gaatcgaaca aaactgcagc
     3601 tgttgctccc atctctgttc ctgctccagt tgctgctgcc gccactgctg ccgccatcac
     3661 tgccactgct gccaccatca ccaccaccat ggtagctgct gctccagttg ctgttgctgc
     3721 tgctgctgct ccagctgctg ctgctgcccc gtctccagcc actgccgctg ctactgctgc
     3781 tgctgtttct ccagctgctg ctggtcagat tccagctgct gcctctgttg cctcagctgc
     3841 tgccgttgct ccttctgctg ctgctgctgc tgctgttcag gttgctccag ctgctccggc
     3901 tccagttcca gctccggctc tggttccggt tccagctcca gcagcggctc aggcttctgc
     3961 tcctgctcag actcaggcac caacttcagc tccggctgtg gctccaactc cagctccaac
     4021 tccaactcca gctgtggctc aggctgaggt tcctgcaagt ccagctaccg gtcctggacc
     4081 acatcgtttg tctataccaa gtttgacctg caatccagac aaaacagatg ggcctgtgtt
     4141 ccactccaat actctggaaa gaaaaactcc cattcagatc ctgggacaag agcctgatgc
     4201 agagatggtg gaatatctca tctaactcga gcatgcatct agagggccct attctatagt
     4261 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     4321 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4381 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4441 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4501 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4561 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4621 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4681 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc
     4741 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4801 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4861 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4921 atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4981 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5041 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5101 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5161 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5221 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5281 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5341 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5401 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5461 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5521 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5581 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5641 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5701 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5761 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5821 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5881 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5941 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6001 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6061 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6121 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6181 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6241 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6301 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6361 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6421 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6481 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6541 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6601 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6661 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6721 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6781 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6841 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6901 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6961 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7021 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7081 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7141 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7201 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7261 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7321 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7381 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7441 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7501 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7621 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7681 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7741 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7801 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7861 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7921 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7981 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8041 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8101 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8161 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8221 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8281 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8341 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8401 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8461 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8521 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8581 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8641 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8701 c
//
