LOCUS       sequence-175223-        5639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express FNTA with a Venus tag at C terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5639)
  AUTHORS   Xu Y, Anderson DE, Ye Y
  TITLE     The HECT domain ubiquitin ligase HUWE1 targets unassembled soluble 
            proteins for degradation.
  JOURNAL   Cell Discov. 2016 Nov 8;2:16040. eCollection 2016.
  PUBMED    27867533
REFERENCE   2  (bases 1 to 5639)
  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 
            Discov."; date: "2016-11-8"; volume: "2"; pages: "16040. eCollection
            2016"
COMMENT     Sequence Label: pVENUS-N1-FNTA
FEATURES             Location/Qualifiers
     source          1..5639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        773..1076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1077..1280
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1231..1251
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1344..1353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2291..2300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2297..2299,2300..2302,2303..3016)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(2342..2363)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2603..2622)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3139..3260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3176..3195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3230..3249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3267..3722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3616..3635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3749..3853
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3855..4212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3855..4046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3878..3898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4063..4198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4247..5041
                     /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             4247..5041
                     /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(4301..4320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4911..4930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5229..5248)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5273..5320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
      721 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      781 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      841 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      901 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      961 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1021 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1081 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1141 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1201 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     1261 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
     1321 agatctcgag ctcaagctta ccgccatggc ggccaccgag ggggtcgggg aggctgcgca
     1381 agggggcgag cccgggcagc cggcgcaacc cccgccccag ccgcacccac cgccgcccca
     1441 gcagcagcac aaggaagaga tggcggccga ggctggggaa gccgtggcgt cccccatgga
     1501 cgacgggttt gtgagcctgg actcgccctc ctatgtcctg tacaggcatt tccggagagt
     1561 tcttttgaag tcacttcaga aggatctaca tgaggaaatg aactacatca ctgcaataat
     1621 tgaggagcag cccaaaaact atcaagtttg gcatcatagg cgagtattag tggaatggct
     1681 aagagatcca tctcaggagc ttgaatttat tgctgatatt cttaatcagg atgcaaagaa
     1741 ttatcatgcc tggcagcatc gacaatgggt tattcaggaa tttaaacttt gggataatga
     1801 gctgcagtat gtggaccaac ttctgaaaga ggatgtgaga aataactctg tctggaacca
     1861 aagatacttc gttatttcta acaccactgg ctacaatgat cgtgctgtat tggagagaga
     1921 agtccaatac actctggaaa tgattaaact agtaccacat aatgaaagtg catggaacta
     1981 tttgaaaggg attttgcagg atcgtggtct ttccaaatat cctaatctgt taaatcaatt
     2041 acttgattta caaccaagtc atagttcccc ctacctaatt gcctttcttg tggatatcta
     2101 tgaagacatg ctagaaaatc agtgtgacaa taaggaagac attcttaata aagcattaga
     2161 gttatgtgaa atcctagcta aagaaaagga cactataaga aaggaatatt ggagatacat
     2221 tggaagatcc cttcaaagca aacacagcac agaaaatgac tcaccaacaa atgtacagca
     2281 accaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2341 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2401 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
     2461 cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta
     2521 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2581 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2641 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2701 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
     2761 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     2821 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2881 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
     2941 gcgcgatcac atggtcctgc tggaacgcgt gaccgccgcc gggatcactc tcggcatgga
     3001 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3061 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3121 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3181 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3241 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3721 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3781 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3841 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4141 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4201 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4261 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4321 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4381 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4441 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4501 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4561 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4621 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4681 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4741 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4801 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4861 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4921 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4981 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5041 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5101 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5161 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5221 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5281 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5341 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5401 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5461 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5521 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5581 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaa
//
