LOCUS       sequence-102369-        8631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pEF1a-puro-T2A-ATHmmuRPL10A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8631)
  AUTHORS   Jan CH, Williams CC, Weissman JS
  TITLE     Principles of ER cotranslational translocation revealed by 
            proximity-specific ribosome profiling.
  JOURNAL   Science. 2014 Nov 7;346(6210):1257521. doi: 10.1126/science.1257521.
            Epub 2014 Nov 6.
  PUBMED    25378630
REFERENCE   2  (bases 1 to 8631)
  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: "Science."; 
            date: "2014-11-7"; volume: "346(6210):1257521. doi"; pages: " 
            10.1126/science.1257521. Epub 2014 Nov 6"
COMMENT     Sequence Label: pJW1511
FEATURES             Location/Qualifiers
     source          1..8631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             844..1024
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1695..1928
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2113..2157
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2455..2572
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2675..2886
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2887..3825
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3782..3802
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3846..4442
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3846..3865)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4342..4362
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4452..4505
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4518..4562
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4575..4595
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4602..4628
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     ncRNA           5359..5373
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    5520..6108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5573..5593)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5991..6002)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6111..6127)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6112..6128)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6637..6817
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6879..7467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6959..6978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7638..8429,8430..8498))
                     /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     8261..8280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8499..8603)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct
     1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
     2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctagg gccgcgtgga
     2641 taaccgtatt accgccatgc atgtgcccgt cagtgggcag agcgcacatc gcccacagtc
     2701 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
     2761 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     2821 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     2881 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
     2941 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
     3001 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
     3061 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
     3121 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
     3181 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
     3241 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
     3301 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
     3361 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     3421 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     3481 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     3541 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     3601 tccaggcacc tcgattagtt ctcgcgcttt tggagtacgt cgtctttagg ttggggggag
     3661 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     3721 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     3781 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgagctagc
     3841 ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     3901 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     3961 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4021 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4081 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4141 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4201 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4261 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4321 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4381 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4441 ccggatcggg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     4501 gcccaccggt cgccaccgga ctgaacgaca tcttcgaggc tcagaaaatc gaatggcacg
     4561 aaggcggagg ttcagaaaat ttgtattttc aaggatcagg ttacccatac gacgtcccag
     4621 actacgctgg atcaagcagc aaagtctctc gcgacaccct gtacgaggcg gtgcgggaag
     4681 tcctgcacgg gaaccagcgc aagcgccgca agtttctgga gacggtggag ctgcagatca
     4741 gcctgaagaa ctacgaccct cagaaggaca aacgtttctc gggcaccgtc aggctcaagt
     4801 ccaccccacg ccccaagttc tcggtgtgcg ttctggggga ccagcagcac tgtgatgaag
     4861 ccaaggccgt ggatatcccc cacatggaca tcgaggcgct caagaagctt aacaaaaaca
     4921 agaagttggt caagaagctg gctaagaagt acgatgcctt tttggcctct gagtctctga
     4981 ttaagcagat cccacgtatc ctgggcccag gcctaaacaa ggctggcaag ttcccctccc
     5041 tgctgacaca caatgaaaac atggtggcca aagtggatga ggtgaaatcg acaatcaagt
     5101 tccagatgaa gaaggtgctg tgtttggccg tcgctgttgg ccacgtgaag atgaccgatg
     5161 atgagctagt ctacaacatt catctggctg tcaatttctt ggtgtccttg cttaagaaaa
     5221 actggcaaaa cgtgcgggct ctgtacatca agagcaccat gggcaagccc cagcgcctat
     5281 attaagaatt gcctagggac tcgagaagaa ggtatattgc tgttgaccca gagcgttggg
     5341 caggtctgaa gttcattagt gaagccacag atgtaatgaa cttcaggacc tgcccactgc
     5401 cttgggcctc ggaattcaag gggtcagtca tctagagttg aaccgactag taagccaggt
     5461 cgagacatgt ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata
     5521 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5581 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5641 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5701 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5761 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5821 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5881 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5941 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6001 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6061 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     6121 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6181 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     6241 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6301 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     6361 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     6421 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     6481 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     6541 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     6601 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     6661 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6721 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6781 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     6841 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6901 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6961 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7021 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7081 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7141 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7201 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7261 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7321 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7381 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7441 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7501 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7561 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7621 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7681 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7741 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7801 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7861 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7921 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7981 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8041 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8101 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8161 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8221 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8281 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8341 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8401 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8461 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8521 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8581 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
