LOCUS       sequence-372334-        7737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express His-tagged NEDD4L FL in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7737)
  AUTHORS   Wright KM, Nathan S, Jiang H, Xia W, Kim H, Chakouri N, Nwafor JN, 
            Fossier L, Srinivasan L, Chen Z, Boronina T, Post J, Paul S, Cole 
            RN, Ben-Johny M, Cole PA, Gabelli SB
  TITLE     NEDD4L intramolecular interactions regulate its auto and substrate 
            Na(V)1.5 ubiquitination.
  JOURNAL   J Biol Chem. 2024 Mar;300(3):105715. doi: 10.1016/j.jbc.2024.105715.
            Epub 2024 Feb 2.
  PUBMED    38309503
REFERENCE   2  (bases 1 to 7737)
  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."; date: "2024-03"; volume: "300(3):105715. doi"; pages: " 
            10.1016/j.jbc.2024.105715. Epub 2024 Feb 2"
COMMENT     Sequence Label: pFastBacHTA-NEDD4L_FL
FEATURES             Location/Qualifiers
     source          1..7737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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     complement(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4101..4121
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    7227..7361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7264..7283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7318..7337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7390..7555
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatcgggcg cggatctcgg tccgaaacca tgtcgtacta ccatcaccat caccatcacg
     4081 attacgatat cccaacgacc gaaaacctgt attttcaggg catggcgacc gggctcgggg
     4141 agccggtcta tggactttcc gaagacgagg gagagtcccg tattctcaga gtaaaagttg
     4201 tttctggaat tgatctcgcc aaaaaggaca tctttggagc cagtgatccg tatgtgaaac
     4261 tttcattgta cgtagcggat gagaatagag aacttgcttt ggtccagaca aaaacaatta
     4321 aaaagacact gaacccaaaa tggaatgaag aattttattt cagggtaaac ccatctaatc
     4381 acagactcct atttgaagta tttgacgaaa atagactgac acgagacgac ttcctgggcc
     4441 aggtggacgt gccccttagt caccttccga cagaagatcc aaccatggag cgaccctata
     4501 catttaagga ctttctcctc agaccaagaa gtcataagtc tcgagttaag ggatttttgc
     4561 gattgaaaat ggcctatatg ccaaaaaatg gaggtcaaga tgaagaaaac agtgaccaga
     4621 gggatgacat ggagcatgga tgggaagttg ttgactcaaa tgactcggct tctcagcacc
     4681 aagaggaact tcctcctcct cctctgcctc ccgggtggga agaaaaagtg gacaatttag
     4741 gccgaactta ctatgtcaac cacaacaacc ggaccactca gtggcacaga ccaagcctga
     4801 tggacgtgtc ctcggagtcg gacaataaca tcagacagat caaccaggag gcagcacacc
     4861 ggcgcttccg ctcccgcagg cacatcagcg aagacttgga gcccgagccc tcggagggcg
     4921 gggatgtccc cgagccttgg gagaccattt cagaggaagt gaatatcgct ggagactctc
     4981 tcggtctggc tctgccccca ccaccggcct ccccaggatc tcggaccagc cctcaggagc
     5041 tgtcagagga actaagcaga aggcttcaga tcactccaga ctccaatggg gaacagttca
     5101 gctctttgat tcaaagagaa ccctcctcaa ggttgaggtc atgcagtgtc accgacgcag
     5161 ttgcagaaca gggccatcta ccaccgccca gtgccccagc tgggagagcg cgttcatcaa
     5221 ctgtcacggg tggtgaggaa ccaacgccat cagtggccta tgtacatact acgccgggtc
     5281 tgccttcagg ctgggaagaa agaaaagatg ctaaggggcg cacatactat gtcaatcata
     5341 acaatcgaac cacaacttgg actcgaccta tcatgcagct tgcagaagat ggtgcgtccg
     5401 gatcagccac aaacagtaac aaccatctaa tcgagcctca gatccgccgg cctcgtagcc
     5461 tcagctcgcc aacagtaact ttatctgccc cgctggaggg tgccaaggac tcacccgtac
     5521 gtcgggctgt gaaagacacc ctttccaacc cacagtcccc acagccatca ccttacaact
     5581 cccccaaacc acaacacaaa gtcacacaga gcttcttgcc acccggctgg gaaatgagga
     5641 tagcgccaaa cggccggccc ttcttcattg atcataacac aaagactaca acctgggaag
     5701 atccacgttt gaaatttcca gtacatatgc ggtcaaagac atctttaaac cccaatgacc
     5761 ttggccccct tcctcctggc tgggaagaaa gaattcactt ggatggccga acgttttata
     5821 ttgatcataa tagcaaaatt actcagtggg aagacccaag actgcagaac ccagctatta
     5881 ctggtccggc tgtcccttac tccagagaat ttaagcagaa atatgactac ttcaggaaga
     5941 aattaaagaa acctgctgat atccccaata ggtttgaaat gaaacttcac agaaataaca
     6001 tatttgaaga gtcctatcgg agaattatgt ccgtgaaaag accagatgtc ctaaaagcta
     6061 gactgtggat tgagtttgaa tcagagaaag gtcttgacta tgggggtgtg gccagagaat
     6121 ggttcttctt actgtccaaa gagatgttca acccctacta cggcctcttt gagtactctg
     6181 ccacggacaa ctacaccctt cagatcaacc ctaattcagg cctctgtaat gaggatcatt
     6241 tgtcctactt cacttttatt ggaagagttg ctggtctggc cgtatttcat gggaagctct
     6301 tagatggttt cttcattaga ccattttaca agatgatgtt gggaaagcag ataaccctga
     6361 atgacatgga atctgtggat agtgaatatt acaactcttt gaaatggatc ctggagaatg
     6421 accctactga gctggacctc atgttctgca tagacgaaga aaactttgga cagacatatc
     6481 aagtggattt gaagcccaat gggtcagaaa taatggtcac aaatgaaaac aaaagggaat
     6541 atatcgactt agtcatccag tggagatttg tgaacagggt ccagaagcag atgaacgcct
     6601 tcttggaggg attcacagaa ctacttccta ttgatttgat taaaattttt gatgaaaatg
     6661 agctggagtt gctcatgtgc ggcctcggtg atgtggatgt gaatgactgg agacagcatt
     6721 ctatttacaa gaacggctac tgcccaaacc accccgtcat tcagtggttc tggaaggctg
     6781 tgctactcat ggacgccgaa aagcgtatcc ggttactgca gtttgtcaca gggacatcgc
     6841 gagtacctat gaatggattt gccgaacttt atggttccaa tggtcctcag ctatttacaa
     6901 tagagcaatg gggcagtcct gagaaactgc ccagagctca cacatgcttt aatcgccttg
     6961 acttacctcc atatgaaacc tttgaagatt tacgagagaa acttctcatg gccgtggaaa
     7021 atgctcaagg atttgaaggg gtggattaag cggccgcttt cgaatctaga gcctgcagtc
     7081 tcgaggcatg cggtaccaag cttgtcgaga agtactagag gatcataatc agccatacca
     7141 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     7201 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     7261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7321 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     7381 ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc
     7441 gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat
     7501 ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg
     7561 gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg
     7621 tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta
     7681 ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg cgaatgg
//
