LOCUS       sequence-328852-        5092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses NEDD4L WW3 in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5092)
  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 5092)
  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: pGEX-PP-NEDD4L-WW3(497-530)
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(549..554,555..572,573..579)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    587..603
                     /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     611..627
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     611..627
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             623..1150
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(643..660)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(643..659)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(652..674)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1252..1257,1258..1273,1274..1280)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1288..1304
                     /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)."
     CDS             1327..1980
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1938..1960
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1987..2004
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2008..2031
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2211..2233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2378..2396)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2464..2568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2569..2637,2638..3429)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2787..2806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3600..4188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4089..4108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4342..4359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4432..4509
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4510..5092,1..500)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4529..4548)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
       61 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
      121 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
      181 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
      241 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
      301 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
      361 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
      421 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
      481 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
      541 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      601 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
      661 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
      721 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      781 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
      841 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
      901 gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc
      961 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     1021 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac
     1081 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
     1141 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
     1201 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt
     1261 aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca
     1321 gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     1381 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     1441 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     1501 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     1561 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     1621 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     1681 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     1741 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     1801 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     1861 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     1921 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     1981 tcggatctgg ttccgcgtgg atctccgctt gaagtcctct ttcagggacc ccatatgagc
     2041 ttcttgccac ccggctggga aatgaggata gcgccaaacg gccggccctt cttcattgat
     2101 cataacacaa agactacaac ctgggaagat ccacgtttgt aaggatcctc gagaattcat
     2161 cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2221 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2281 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     2341 agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     2401 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3001 gagctaaccg cttttttgca caacatggga gatcatgtaa ctcgccttga tcgttgggaa
     3061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     3121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3721 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4261 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4321 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4381 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc
     4441 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     4501 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     4561 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     4621 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     4681 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     4741 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     4801 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     4861 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     4921 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     4981 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     5041 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gc
//
