LOCUS       sequence-408401-        8940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human NEDD4L (delta C2 isoform).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8940)
  AUTHORS   Horns F, Martinez JA, Fan C, Haque M, Linton JM, Tobin V, Santat L, 
            Maggiolo AO, Bjorkman PJ, Lois C, Elowitz MB
  TITLE     Engineering RNA export for measurement and manipulation of living 
            cells.
  JOURNAL   Cell. 2023 Jul 10:S0092-8674(23)00689-X. doi: 
            10.1016/j.cell.2023.06.013.
  PUBMED    37437570
REFERENCE   2  (bases 1 to 8940)
  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: "Cell. 2023 
            Jul 10:S0092-8674(23)00689-X. doi: 10.1016/j.cell.2023.06.013."
COMMENT     Sequence Label: pJAM1.20_PB-NEDD4L_deltaC2-PGK-Blast
FEATURES             Location/Qualifiers
     source          1..8940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   14..48
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        660..871
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             884..1152
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3878..3999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3915..3934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3969..3988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4372..4391
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      4469..4478
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4475..4873
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4891..5115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5542..5604
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6103..6119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6103..6119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6116..6138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6127..6143
                     /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(6151..6157,6158..6175,6176..6181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6196..6217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6334..6351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6505..7093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6585..6604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7264..8058)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(7264..8058)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7375..7394)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7985..8004
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(8059..8163)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8189..8644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8326..8347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8538..8557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8771..8793
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8785..8802
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8786..8802
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcagctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
       61 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      121 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      181 ataacaaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      241 actcatacga taattatatt gttatttcgt gttctactta cgtgataact tattatatat
      301 atattttctt gttatagata tccttgaggg acagcccccc cccaaagccc ccagggatgt
      361 aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc
      421 ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg
      481 tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg
      541 gggggatacg gggaaaaatc gattaactat aacggtccta aggtagcgaa cacctgcgct
      601 cgagcgacgt cattaccctg ttatccctag gatctgcgat cgctccggtg cccgtcagtg
      661 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      721 gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      781 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      841 ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc
      901 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc
      961 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag
     1021 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc
     1081 cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg
     1141 ccgttacaga tccaagctgt gaccggcgcc tacgctagcg ccaccatgga gcgaccctat
     1201 acatttaagg actttctcct cagaccaaga agtcataagt ctcgagttaa gggatttttg
     1261 cgattgaaaa tggcctatat gccaaaaaat ggaggtcaag atgaagaaaa cagtgaccag
     1321 agggatgaca tggagcatgg atgggaagtt gttgactcaa atgactcggc ttctcagcac
     1381 caagaggaac ttcctcctcc tcctctgcct cccgggtggg aagaaaaagt ggacaattta
     1441 ggccgaactt actatgtcaa ccacaacaac cggaccactc agtggcacag accaagcctg
     1501 atggacgtgt cctcggagtc ggacaataac atcagacaga tcaaccagga ggcagcacac
     1561 cggcgcttcc gctcccgcag gcacatcagc gaagacttgg agcccgagcc ctcggagggc
     1621 ggggatgtcc ccgagccttg ggagaccatt tcagaggaag tgaatatcgc tggagactct
     1681 ctcggtctgg ctctgccccc accaccggcc tccccaggat ctcggaccag ccctcaggag
     1741 ctgtcagagg aactaagcag aaggcttcag atcactccag actccaatgg ggaacagttc
     1801 agctctttga ttcaaagaga accctcctca aggttgaggt catgcagtgt caccgacgca
     1861 gttgcagaac agggccatct accaccgccc agtgccccag ctgggagagc gcgttcatca
     1921 actgtcacgg gtggtgagga accaacgcca tcagtggcct atgtacatac cacgccgggt
     1981 ctgccttcag gctgggaaga aagaaaagat gctaaggggc gcacatacta tgtcaatcat
     2041 aacaatcgaa ccacaacttg gactcgacct atcatgcagc ttgcagaaga tggtgcgtcc
     2101 ggatcagcca caaacagtaa caaccatcta atcgagcctc agatccgccg gcctcgtagc
     2161 ctcagctcgc caacagtaac tttatctgcc ccgctggagg gtgccaagga ctcacccgta
     2221 cgtcgggctg tgaaagacac cctttccaac ccacagtccc cacagccatc accttacaac
     2281 tcccccaaac cacaacacaa agtcacacag agcttcttgc cacccggctg ggaaatgagg
     2341 atagcgccaa acggccggcc cttcttcatt gatcataaca caaagactac aacctgggaa
     2401 gatccacgtt tgaaatttcc agtacatatg cggtcaaaga catctttaaa ccccaatgac
     2461 cttggccccc ttcctcctgg ctgggaagaa agaattcact tggatggccg aacgttttat
     2521 attgatcata atagcaaaat tactcagtgg gaagacccaa gactgcagaa cccagctatt
     2581 actggtccgg ctgtccctta ctccagagaa tttaagcaga aatatgacta cttcaggaag
     2641 aaattaaaga aacctgctga tatccccaat aggtttgaaa tgaaacttca cagaaataac
     2701 atatttgaag agtcctatcg gagaattatg tccgtgaaaa gaccagatgt cctaaaagct
     2761 agactgtgga ttgagtttga atcagagaaa ggtcttgact atgggggtgt ggccagagaa
     2821 tggttcttct tactgtccaa agagatgttc aacccctact acggcctctt tgagtactct
     2881 gccacggaca actacaccct tcagatcaac cctaattcag gcctctgtaa tgaggatcat
     2941 ttgtcctact tcacttttat tggaagagtt gctggtctgg ccgtatttca tgggaagctc
     3001 ttagatggtt tcttcattag accattttac aagatgatgt tgggaaagca gataaccctg
     3061 aatgacatgg aatctgtgga tagtgaatat tacaactctt tgaaatggat cctggagaat
     3121 gaccctactg agctggacct catgttctgc atagacgaag aaaactttgg acagacatat
     3181 caagtggatt tgaagcccaa tgggtcagaa ataatggtca caaatgaaaa caaaagggaa
     3241 tatatcgact tagtcatcca gtggagattt gtgaacaggg tccagaagca gatgaacgcc
     3301 ttcttggagg gattcacaga actacttcct attgatttga ttaaaatttt tgatgaaaat
     3361 gagctggagt tgctcatgtg cggcctcggt gatgtggatg tgaatgactg gagacagcat
     3421 tctatttaca agaacggcta ctgcccaaac caccccgtca ttcagtggtt ctggaaggct
     3481 gtgctactca tggacgccga aaagcgtatc cggttactgc agtttgtcac agggacatcg
     3541 cgagtaccta tgaatggatt tgccgaactt tatggttcca atggtcctca gctgtttaca
     3601 atagagcaat ggggcagtcc tgagaaactg cccagagctc acacatgctt taatcgcctt
     3661 gacttacctc catatgaaac ctttgaagat ttacgagaga aacttctcat ggccgtggaa
     3721 aatgctcaag gatttgaagg ggtggattag gcggccgcgt cgacgactct agatcataat
     3781 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3841 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3901 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3961 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa gcgccggcgc tacacaagtg
     4021 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     4081 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     4141 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     4201 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     4261 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     4321 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     4381 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     4441 cctgcagccc aagcttacca tgggtaccgc caccatggcc aagcctttgt ctcaagaaga
     4501 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagatta
     4561 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     4621 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     4681 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     4741 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     4801 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     4861 gtgggagggc taaggtcacc cgcgcgcgga ctgtgccttc tagttgccag ccatctgttg
     4921 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4981 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     5041 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     5101 cggtgggctc tatggttcga accagtacga tggttttccc cgtatccccc caggtgtctg
     5161 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt
     5221 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc
     5281 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg
     5341 gggctttggg ggggggctgt ccctgataaa agttttgtta ctttatagaa gaaattttga
     5401 gtttttgttt ttttttaata aataaataaa cataaataaa ttgtttgttg aatttattat
     5461 tagtatgtaa gtgtaaatat aataaaactt aatatctatt caaattaata aataaacctc
     5521 gatatacaga ccgataaaac acatgcgtca attttacgca tgattatctt taacgtacgt
     5581 cacaatatga ttatctttct agggttaatc tagctgcggc gcgccatcag gatcatatcg
     5641 tcgggtcttt tttccggctc agtcatcgcc caagctggcg ctatctgggc atcggggagg
     5701 aagaagcccg tgccttttcc cgcgaggttg aagcggcatg gaaagagttt gccgaggatg
     5761 actgctgctg cattgacgtt gagcgaaaac gcacgtttac catgatgatt cgggaaggtg
     5821 tggccatgca cgcctttaac ggtgaactgt tcgttcaggc cacctgggat accagttcgt
     5881 cgcggctttt ccggacacag ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca
     5941 ataccggcga cagccggaac tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc
     6001 tgggatatta cgtcagcgag gacgggtatc ctggctggat gccgcagaaa tggacatgga
     6061 taccccgtga gttacccggc gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     6121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7261 cagtcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     7321 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     7381 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     7441 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     7501 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     7561 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     7621 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     7681 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     7741 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     7801 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     7861 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     7921 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     7981 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     8041 tccatcttgt tcaatcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8161 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     8221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     8281 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     8341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     8401 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     8461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     8521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     8581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     8641 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     8701 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     8761 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc ctcgttcatt
     8821 cacgtttttg aacccgtgga ggacgggcag actcgcggtg caaatgtgtt ttacagcgtg
     8881 atggagcaga tgaagatgct cgacacgctg cagaacacga gctccaccgc ggggcgcgcc
//
