LOCUS       sequence-338240-        8687 bp    DNA     circular SYN 15-APR-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 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(E104K/E105K)
FEATURES             Location/Qualifiers
     source          1..8687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(132..151)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        323..702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        703..906
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     857..877
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     951..970
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        951..969
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             992..1018
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1022..1048
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2207..2227
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             3757..3774
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(4328..4346)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4329..4346)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    4372..4596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4642..5070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4729..4748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4939..4960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5079..5099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5084..5413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5264..5399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5326..5345
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5480..6274
                     /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(5534..5553)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6144..6163
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6448..6569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6485..6504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6539..6558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6618..6634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6618..6634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6631..6653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6642..6658
                     /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(6666..6672,6673..6690,6691..6696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6711..6732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6849..6866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7020..7608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7100..7119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7779..8570,8571..8639))
                     /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     8402..8421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8640..8687,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      121 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      181 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      241 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      301 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      361 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      421 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      481 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      601 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      721 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      781 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      841 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      901 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      961 cactataggg agacccaagc ttgccatggc ctacccatac gatgttcctg actatgcggg
     1021 ctatccctat gacgtcccgg actatgcagg atccatgaga aattctgacg aacagccaag
     1081 tggagggacc acggtattgc agcgtttgct acaagagcag cttcgctatg gcaatcctag
     1141 tgagaatcgc agcctgcttg ccatacacca gcaagccaca gggaatggcc ctcctttccc
     1201 cagtggcagt gggaacccgg gccctcagag tgatgtgttg agtccccaag accaccacca
     1261 acagcttgtg gctcatgctg ctcgacaaga accccagggg caggaaatcc agtcagaaaa
     1321 cctcatcatg gagaagcagc tgtctcctcg aatgcaaaat aataaaaaac tcccgaccta
     1381 tgaagaagcc aaggtccagt cccagtactt tcggggccaa cagcatgcca gtgttggagc
     1441 tgccttctat gtcactggag tcaccaacca gaagatgagg actgagggac gcccatcagt
     1501 tcagcggctc aatcctggaa agatgcacca agatgaggga ctcagagacc ttaagcaagg
     1561 gcatgtccgt tccttgagtg aacgactaat gcagatgtca ctggccacca gtggagttaa
     1621 ggcccatcca cctgttacca gtgctcccct ctccccacca caacccaatg acctctacaa
     1681 gaatcccaca agttccagtg aattctacaa ggcccaaggg ccacttccta accagcatag
     1741 cctgaagggc atggaacacc gaggcccccc accagaatat cccttcaagg gcatgccacc
     1801 ccaatctgta gtgtgcaagc cccaagagcc agggcacttc tatagtgagc atcgcctgaa
     1861 ccagccaggg agaacagagg ggcaactgat gaggtatcag catccccctg agtatggagc
     1921 agccaggcca gcgcaggaca tctcattgcc attgtcagcc aggaactcgc agcctcacag
     1981 ccctacttct tctctgacct ctggggggtc cttgcccttg ctacaatctc caccatccac
     2041 tagattgtct cctgcccgac accccttggt cccaaaccag ggagaccatt cagctcacct
     2101 gcctaggccg cagcagcatt tccttcctaa tcaggctcac cagggggatc attaccgtct
     2161 ctcccaacct ggcctgagtc agcagcagca gcaacagcag cagcagcacc atcatcacca
     2221 tcaccaccaa caacagcagc agcagcagcc acagcagcag ccaggagaag cctattcagc
     2281 tatgcctcgg gctcagccat cctctgcttc ttatcagcca gtgccagcag acccttttgc
     2341 cattgtttcc agagcccagc agatggttga gatcctctca gacgagaacc ggaacttgag
     2401 gcaagagttg gaaggatgct atgagaaggt ggcaagactg cagaaggtgg agacagaaat
     2461 ccagcgcgtc tcggaggcat atgagaacct cgtgaagtca tcctccaaaa gagaggccct
     2521 agagaaagcc atgagaaaca agctagaggg cgagattcgg aggatgcatg atttcaacag
     2581 ggatctgaga gagcgtctag agactgccaa caagcagctt gcagagaagg aatatgaggg
     2641 gtcagaggac accagaaaaa ccatctcgca gctctttgca aaaaataaag aaagccagcg
     2701 tgagaaggag aagctggaag cggagctggc cactgcccgt tctaccaatg aggaccaaag
     2761 acgacacatc gaaatccgag atcaggccct gagtaatgcc caggccaagg tggtaaagct
     2821 ggaagaagag ctgaaaaaga agcaagtgta cgttgacaag gtggagaaga tgcagcaggc
     2881 ccttgtacag ctccaggcag catgtgaaaa acgtgagcag ctagagcacc gtctccggac
     2941 acgactggag agggaactgg aatccctgag aatccagcag cgtcagggca actgtcagcc
     3001 caccaacgtt tcagaataca atgctgctgc actgatggag ctccttcggg agaaagagga
     3061 gaggattctg gctctggaag ctgatatgac aaagtgggag cagaaatatt tggaggagaa
     3121 tgtgatgaga cattttgctc tggatgctgc tgcaactgtg gctgctcaga gggacacaac
     3181 agtcatcagt cactctccta acaccagcta tgacacagct ctagaagctc gcatccagaa
     3241 agaggaggaa gaaatcttga tggccaataa gcgttgcctt gacatggagg gcaggattaa
     3301 gaccctccat gcccagatta ttgagaagga tgccatgatc aaagtactcc agcagcgttc
     3361 ccggaaggag ccgagcaaga cagagcagct gtcgtgcatg cggccagcga agtctctgat
     3421 gtccatttcc aatgctggat caggcttgct ctcccactca tccaccctga ctggctcccc
     3481 catcatggaa gaaaagcgag acgacaagag ctggaagggg agcctaggca ttctcctggg
     3541 tggagactac cgtgctgaat atgtcccttc cacaccctcg cctgtgccac cctcgactcc
     3601 cctgctctcg gctcactcca agacagacag ccgagactgc agtacccaaa ctgaacgtgg
     3661 gacggaatcg aacaaaactg cagctgttgc tcccatctct gttcctgctc cagttgctgc
     3721 tgccgccact gctgccgcca tcactgccac tgctgccacc atcaccacca ccatggtagc
     3781 tgctgctcca gttgctgttg ctgctgctgc tgctccagct gctgctgctg ccccgtctcc
     3841 agccactgcc gctgctactg ctgctgctgt ttctccagct gctgctggtc agattccagc
     3901 tgctgcctct gttgcctcag ctgctgccgt tgctccttct gctgctgctg ctgctgctgt
     3961 tcaggttgct ccagctgctc cggctccagt tccagctccg gctctggttc cggtcccagc
     4021 tccagcagcg gctcaggctt ctgctcctgc tcagactcag gcaccaactt cagctccggc
     4081 tgtggctcca actccagctc caactccaac tccagctgtg gctcaggctg aggttcctgc
     4141 aagtccagct accggtcctg gaccacatcg tttgtctata ccaagtttga cctgcaatcc
     4201 agacaaaaca gatgggcctg tgttccactc caatactctg gaaagaaaaa ctcccattca
     4261 gatcctggga caagagcctg atgcagagat ggtggaatat ctcatctaac tcgagggggg
     4321 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     4381 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4441 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4501 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4561 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4621 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4681 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4981 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5041 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     5101 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5161 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5221 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5281 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5341 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5401 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     5461 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5521 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5581 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5641 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5701 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5761 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5821 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5881 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5941 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6001 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6061 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6121 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6181 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6241 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     6301 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6361 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6421 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6481 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6541 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6601 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6661 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6721 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6781 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6841 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6901 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6961 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7021 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7081 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7141 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7201 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7261 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7321 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7381 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7441 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7501 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7561 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7621 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7681 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7741 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7801 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7861 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7921 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7981 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8041 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8101 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8161 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8221 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8281 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8341 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8401 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8461 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8521 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8581 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8641 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtct
//
