LOCUS       sequence-483846-        6607 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Mammalian expression of HyPerFLEX targeted to the cytosolic side of 
            the endoplasmic reticulum membranes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6607)
  TITLE     Belousov lab plasmids 2025
REFERENCE   2  (bases 1 to 6607)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-CMV-HyPerFLEX-Sec61b
FEATURES             Location/Qualifiers
     source          1..6607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     710..734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..1240
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             2431..2721
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     misc_feature    2734..3322
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2787..2807)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3205..3216)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3354..3830
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3553..3572)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3870..4010
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3870..3999
                     /label=AAV2 ITR
     rep_origin      4085..4540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4382..4403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4557..4576)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4676..4698
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4736..4754)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4822..4926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4927..4995,4996..5787)
                     /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     complement(5145..5164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5958..6546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6447..6466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat tggaacgcgg
      841 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacaaaaaat
      901 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
      961 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     1021 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     1081 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     1141 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     1201 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     1261 ctgtgtgctg gcccatcact ttggcaaaga attgggattc gaacatcgat tgaattctga
     1321 gccaccatgc acctggctaa tgaggagcaa aacgaattgg aaggtgcttt taagctgggc
     1381 ttgatcttta ccgtcgcccc gtacttgctg cctaagttga tcgtcagcct gcgccgcacc
     1441 gccccgaaga tgcctttgat gttggaggag aattacaccc acacccttac cgagtcactg
     1501 aaacgcggcg acgttgacgc catcattgtg gcagagccgt ttcaagagcc gggtattgtg
     1561 actgaacctt tgtatgacga gccgtttttc gttatcgttc cgaagggcca tagcttcgag
     1621 gagttggatg ccgttagccc tcgcatgttg ggcgaagaac aagttctgct tctgaccgag
     1681 ggcaactgta tgcgtgacca ggtgcttagc atttttacca aggtcaaggt gcacatgaag
     1741 aaagcccttt ccggtgacag ctattgggtc tttgtgaaac gggtaggtgg cactggcggt
     1801 gagcatgttg tctttggcag tgaggacatc gagaacacta tggccaaaat ggacgacgga
     1861 caactggatg ggttggcctt tggcgcaatt cagctcgatg gtgacgggaa tatcctgcag
     1921 tacaatgctg ctgaaggaga catcacaggc agagatccca aacaggtgat tgggaagaac
     1981 ttcttcaagg atgttgcacc tggaacggat tctcccgagt tttacggcaa attcaaggaa
     2041 ggcgtagcgt cagggaatct gaacaccatg ttcgaatgga tgataccgac tcttaccgat
     2101 catagttgca gcgagcttgc cgccaaacag cgcattcagg gtctcaccaa cacccttcaa
     2161 ggctcaagca tcgataccat ccgccacatg gttgccagtg gcttggcaat cagcgtcttg
     2221 cctgccactg cccttaccga aaacgaccac atgctgttta gcattatccc gtttgaaggc
     2281 accccgccta gccgccgcgt tgtcctggcc tatcgccgca actttgtccg ccctaaggcc
     2341 ctttcagcca tgaaagccgc aattatgcaa agccaactgc atggtgttag cttcatctgc
     2401 gataccggta gcggatccgg actcagatct atgcctggtc cgacccccag tggcactaac
     2461 gtgggatcct cagggcgctc tcccagcaaa gcagtggccg cccgggcggc gggatccact
     2521 gtccggcaga ggaaaaatgc cagctgtggg acaaggagtg caggccgcac aacctcggca
     2581 ggcaccgggg ggatgtggcg attctacaca gaagattcac ctgggctcaa agttggccct
     2641 gttccagtat tggttatgag tcttctgttc atcgcttctg tatttatgtt gcacatttgg
     2701 ggcaagtaca ctcgttcgta gaagcttatc gataatcaac ctctggatta caaaatttgt
     2761 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2821 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2881 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2941 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3001 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3061 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3121 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     3181 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3241 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3301 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc
     3361 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     3421 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     3481 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     3541 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     3601 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     3661 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     3721 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     3781 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt
     3841 aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     3901 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3961 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4021 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4081 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4141 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     4201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4381 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4441 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4501 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4561 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4621 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4681 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4741 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4861 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4921 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4981 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5041 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5101 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5161 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5221 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5281 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5341 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5401 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5461 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5521 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5581 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5641 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5701 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5761 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5821 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5881 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5941 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6601 cacatgt
//
