LOCUS       sequence-363112-        7295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of a ER-residential PEN2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7295)
  AUTHORS   Ma T, Tian X, Zhang B, Li M, Wang Y, Yang C, Wu J, Wei X, Qu Q, Yu 
            Y, Long S, Feng JW, Li C, Zhang C, Xie C, Wu Y, Xu Z, Chen J, Yu Y, 
            Huang X, He Y, Yao L, Zhang L, Zhu M, Wang W, Wang ZC, Zhang M, Bao 
            Y, Jia W, Lin SY, Ye Z, Piao HL, Deng X, Zhang CS, Lin SC
  TITLE     Low-dose metformin targets the lysosomal AMPK pathway through PEN2.
  JOURNAL   Nature. 2022 Mar;603(7899):159-165. doi: 10.1038/s41586-022-04431-8.
            Epub 2022 Feb 23.
  PUBMED    35197629
REFERENCE   2  (bases 1 to 7295)
  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"; doi: 
            "10.1038/s41586-022-04431-8"; journalName: "Nature"; date: 
            "2022-03"; volume: "603"; issue: "7899"; pages: "159-165"
COMMENT     Sequence Label: pBoBi-hSEC61B-C-PEN2
FEATURES             Location/Qualifiers
     source          1..7295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     158..177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        293..340
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             359..733
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    863..984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(900..919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     954..973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1033..1049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1033..1049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1046..1068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1057..1073
                     /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(1081..1087,1088..1105,1106..1111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1126..1147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1264..1281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1435..2023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1515..1534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2194..2985,2986..3054))
                     /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     2817..2836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3055..3159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3234..3253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3425..3804
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3805..4008
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3959..3979
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4005..4029
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4116..4403
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     CDS             4725..4751
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4758..4784
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4788..4814
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          4886..5115
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5098..5117
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    5292..5880
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5345..5365)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5763..5774)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6287..6467
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6493..6510)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6499..6723
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6769..7197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6856..6875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7066..7087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7206..7226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7211..7295,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
      301 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
      361 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
      421 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
      481 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
      541 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
      601 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
      661 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
      721 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
      781 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
      841 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
      901 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
      961 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     1021 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1081 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1141 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1201 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1261 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1321 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1381 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1441 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1501 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1561 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1621 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1681 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1741 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1801 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1861 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1921 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1981 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2041 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2101 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2161 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2221 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2281 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2341 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2401 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2461 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2521 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2581 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2641 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2701 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2761 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2821 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2881 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2941 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3001 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3061 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3121 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3181 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     3241 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     3301 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     3361 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     3421 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     3481 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3541 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3601 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3661 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3721 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3781 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     3841 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     3901 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     3961 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     4021 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgactg
     4081 agctccttaa ggttaactct agagaattcg gatccatgcc tggtccgacc cccagtggca
     4141 ctaacgtggg atcctcaggg cgctctccca gcaaagcagt ggccgcccgg gcggcgggat
     4201 ccactgtccg gcagaggaaa aatgccagct gtgggacaag gagtgcaggc cgcacaacct
     4261 cggcaggcac cggggggatg tggcgattct acacagaaga ttcacctggg ctcaaagttg
     4321 gccctgttcc agtattggtt atgagtcttc tgttcatcgc ttctgtattt atgttgcaca
     4381 tttggggcaa gtacactcgt tcgatgaacc tggagcgagt gtccaatgag gagaaattga
     4441 acctgtgccg gaagtactac ctgggggggt ttgctttcct gccttttctc tggttggtca
     4501 acatcttctg gttcttccga gaggccttcc ttgtcccagc ctacacagaa cagagccaaa
     4561 tcaaaggcta tgtctggcgc tcagctgtgg gcttcctctt ctgggtgata gtgctcacct
     4621 cctggatcac catcttccag atctaccggc cccgctgggg tgcccttggg gactacctct
     4681 ccttcaccat acccctgggc acccccctcg agccatggaa gctttaccct tatgacgtac
     4741 ctgactatgc tggcgcctac ccttatgacg tacctgacta tgctggatac ccttatgacg
     4801 tacctgacta tgctggagat atctaactga ctgaaccggt ggtaccgagg aattaattcg
     4861 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     4921 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     4981 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     5041 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     5101 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     5161 cggccaattc cgccgatcct ggaaaaacat ggagcaatca caagtagcaa tacagcagct
     5221 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     5281 acacctcaga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5341 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5401 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5461 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5521 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5581 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5641 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     5701 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5761 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5821 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5881 ctggaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5941 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     6001 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     6061 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     6121 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     6181 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     6241 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     6301 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6361 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6421 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg ccgctttaaa
     6481 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6541 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6601 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6661 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     6721 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     6781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6961 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     7201 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     7261 tatgcaaagc atgcatctca attagtcagc aacca
//
