LOCUS       sequence-10016-e        8645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8645)
  TITLE     PERK constructs
REFERENCE   2  (bases 1 to 8645)
  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"
COMMENT     Sequence Label: PERK 7: PERK.K618A.9E10.pBABE
FEATURES             Location/Qualifiers
     source          1..8645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    523..722
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             923..1339
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1269..1291
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1307..1323
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             4912..4941
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4956..4976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4961..5290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5141..5276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5300..5899
                     /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(5300..5319)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5796..5816
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6120..6589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6719..6736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6890..7478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6970..6989)
                     /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
     LTR             join(8634..8645,1..458)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
       61 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
      121 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
      181 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
      241 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
      301 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
      361 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
      421 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
      481 ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta
      541 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
      601 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      661 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
      721 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
      781 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
      841 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      901 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
      961 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1021 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1081 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1141 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1201 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1261 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1321 cactccttct ctaggcgccg gccggatccc cagtgtggtg gtacgtagaa ttcggcacga
     1381 gcgatgtctg cacaaggctg tcactcaggt ggcagtggct gacacgtggc cgggcagctc
     1441 tgctgctgcg gcgcgaagtc gagaggcggc ggggtccgtg gcgcgcgctc gcattgctcc
     1501 gaggctccga gcggcgatac gggcgggcgc cgacggcagg gtctccatgc ccgcgcgtgg
     1561 ggcgggccgc tgatggagcg cgccacccgg cccgggccgc gcgcgctgct gctgcttctg
     1621 ttcctgctgc tgggctgcgc ggcggggatc tctgcggtcg cgcccgcccg cagtttgctt
     1681 gctcccgcgt cggagacagt gtttggctta ggggcagcgg ccgccccgac ttcggccgcg
     1741 cgggtgcctg cggtggcaac ggccgaagtg accgtggagg acgccgaggc attgccggct
     1801 gccgctggcg aaccggagtc acgcgcgacg gagcccgatg acgacgtgga actgcggcct
     1861 cgcggcaggt ccttggtaat catcagcact ttagatggac gaatcgctgc actggatgcc
     1921 gagaatgatg ggaaaaagca gtgggatttg gacgtggggt ctggttcctt ggtttcatct
     1981 agcctcagca agccagaggt gtttgggaac aagatgatca tcccctccct ggatggagac
     2041 ctcttccagt gggaccggga ccgagagagc atggaggccg tccccttcac ggtggagtcc
     2101 ctgctcgaat cttcctacaa gtttggagat gatgttgttc tggttggagg gaaatctctg
     2161 attacatacg gactcagtgc ttacagtgga aagctgaggt atatctgttc tgccttggga
     2221 tgtcgccgat gggatagtga tgaaatggaa gaagaggaag acatcttgct tctgcagcgt
     2281 acgcagaaga ctgtgcgagc tgtcgggcct cgaagcggca gtgagaagtg gaatttcagt
     2341 gttggccact ttgaacttcg gtatattcca gacatggaaa ctagagccgg attcattgaa
     2401 agcaccttta aaccgggtgg aaacaaagaa gactctaaaa ttatttcaga tgtggaagaa
     2461 caagaagcca ccatgctgga cacagtgata aaagtttccg ttgctgattg gaaggtcatg
     2521 gcgtttagta ggaagggagg ccgcctggaa tgggagtacc agttttgtac tcccatcgcg
     2581 tccgcctggc tggtgaggga tggcaaggtc atccccatca gcctgtttga tgatacaagt
     2641 tacacagcca gcgaagaagc cttgggagac gaagaagaca ttgtagaggc tgctcgggga
     2701 gccacagaga acagcgtgta cttagggatg tacagaggcc agctgtacct gcagtcgtcc
     2761 gtcagggtct cagaaaagtt ccctacaagc ccaaaggcct tggagtctgt aaatggcgaa
     2821 aatgcaatta ttcctctgcc gacgatcaaa tggaagccct taatccattc tccttctagg
     2881 actcctgtct tggttgggtc tgatgaattt gacaaatgtc taagtaatga taagtattcc
     2941 cacgaagaat acagtaatgg tgcactttca atcctccagt atccatacga taacggttac
     3001 tatctgccat actacaagag agaaaggaat aagcggagca cgcagatcac agtcaggttc
     3061 ctggacagcc cccactacag caagaacatc cgcaagaagg accctatcct cctgctgcac
     3121 tggtggaagg agatattcgg gacgatcctg ctttgcatcg tagccacgac cttcatcgtg
     3181 cgcaggcttt tccatcctca gccccacagg cagcggaagg agtctgaaac tcagtgccag
     3241 actgaaagta aatacgactc cgtgagtgcc gatgtcagtg acaacagctg gaatgacatg
     3301 aagtactcag gatacgtatc ccgatatcta acagattttg agccaattca gtgcatgggt
     3361 cgtggtggct ttggcgttgt ctttgaagct aaaaacaaag tagatgactg caattacgct
     3421 atcgcgagga tccggctccc caacagggag ttggcacggg agaaggtaat gcgggaagtt
     3481 aaagccttgg ctaagctgga acacccaggc attgtgaggt atttcaacgc ctggctggaa
     3541 accccaccag agaagtggca agaagagatg gatgagatct ggctcaaaga cgaaagcaca
     3601 gactggccgc tcagctcccc tagcccgatg gatgccccat ctgttaagat ccgaaggatg
     3661 gatccnttct ctacaaaaga gcagatcgaa gtcatagctc cttctcctga aagaagtcgg
     3721 tctttctcgg tgggcatttc ctgtggccag acaagctcat cggagagcca gttctctccc
     3781 ctggagttct cagggacaga ctgcggagac aacagtgact cagcggacgc agcctacaac
     3841 ctccaggaca gttgcctgac ggactgcgag gacgtggaag atggcaccgt ggacggcaat
     3901 gacgagggac actcctttga actttgtccg tccgaagctt ctccctatac ccggtctagg
     3961 gaaggaacgt cctcctccat agtgtttgag gactctggct gcggcaacgc gtccagtaag
     4021 gaggagccca gagggaaccg gctgcatgat ggcaaccatt atgttaataa gctaactgat
     4081 ctcaagtgct ccagcagcag gtcttcttca gaagccacca ccttgtctac ctcccctacc
     4141 aggccaacca ctctaagctt ggatttcacc aagaacactg tgggccagct ccagcccagc
     4201 tcccccaagg tgtatctgta cattcagatg cagctgtgca ggaaggagaa cctcaaagac
     4261 tggatgaacc ggcgctgcag cttggaggac cgggagcacg gcgtgtgcct gcacatcttc
     4321 ctgcagatcg cagaggcagt ggagttcctg cacagcaagg gactcatgca cagggacctc
     4381 aagccttcca acatattctt cacaatggat gatgtggtca aggttgggga ctttggactg
     4441 gtgactgcta tggaccaaga tgaagaagag cagactgttc tgactccaat gccagcctat
     4501 gctacgcaca cgggacaagt agggaccaag ctatacatga gcccagagca gattcatgga
     4561 aacaactact cccataaagt ggacatcttc tctttaggct tgattctgtt tgaactcctc
     4621 tacccattca gcacccagat ggaacgagtc cggattttaa ctgatgtcag aaatctcaag
     4681 tttcctctac tgttcactca gaaatatccc caagagcata tgatggttca agacatgctc
     4741 tctccatccc ccacggagcg gcctgaagcc acagacatca ttgaaaatgc catatttgag
     4801 aacttggagt ttcccgggaa aacggttctg agacagcggt cccgctccat gagttcatct
     4861 ggaacaaaac attccagaca gcccagctgc tcgtacacgc cactgcctgg cgagcagaag
     4921 ctgatctctg aggaggatct gtgactcgac cctgtggaat gtgtgtcagt tagggtgtgg
     4981 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5041 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5101 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5161 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5221 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5281 cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5341 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5401 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5461 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5521 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5581 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5641 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5701 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5761 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5821 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5881 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     5941 acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6001 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6061 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     6121 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     6181 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6241 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     6301 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     6361 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     6421 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     6481 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     6541 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     6601 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     6661 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6721 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6781 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7081 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7321 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7561 caaaaaggat cttcacctag atccttttgc ggccgcaaat 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 tgccacctgc agcctgaata
     8641 tgggc
//
