LOCUS       sequence-10062-e        8219 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 8219)
  TITLE     IRE1 plasmids
REFERENCE   2  (bases 1 to 8219)
  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: IRE1b(K536A).9E10.pCDNA3
FEATURES             Location/Qualifiers
     source          1..8219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3711..3740
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3771..3789)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3772..3789)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3809..3826)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3815..4039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4085..4513
                     /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(4522..4542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4527..4857
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4708..4843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4770..4789
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4924..5718
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4978..4997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5588..5607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5892..6013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5929..5948)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5983..6002
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6062..6078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6062..6078)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6075..6097)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6086..6102
                     /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(6110..6116,6117..6134,6135..6140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6155..6176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6293..6310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6464..7052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6544..6563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7223..8014,8015..8083))
                     /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     7846..7865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8084..8188)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcggcacga ggagactttg
      961 tggttgccca gactatggcg agaccagtcc agagatttca actgtggtct cctctcgggt
     1021 tcctgctgca gcttgtgacg ttgctgggga aactcggtcc acaggtccag tctgtcaggc
     1081 cagagagcct tctgtttgtg tctaccttgg acggaagtct ccatgcactg aacaagcaga
     1141 caggggactt gaagtggaca gtgaaggatg accccatcat ccagggccca atgtatgtca
     1201 cagaaatggc ctttctctct gacccagctg acggcagcct gtatgtcttg ggaacccaga
     1261 aacaacaggg gctgatgaaa ctgccattca ccatcccaga gctggttcat gcctctccct
     1321 gtcgcagctc tgatggtgtc ttctacacag gccggaagca ggatgcctgg tttgtagtgg
     1381 accctgagtc aggggagact cagatgacac tgaccacaga aggcctctct actcctcaac
     1441 ttttcattgg ccgaacacag tacacagtct ccatgcatga cctacggacc ccagccctgc
     1501 ggtggaacac aacctatcgc cgctactcag cacccctcct gaatggctca cccgggaaat
     1561 acatgagcca cctgacatcc tgtgggatgg gcctgcttct cactgtggat ccaggaagtg
     1621 gaattgtgct gtggacacag gacctgggag tgcctgtgac agggatttac acatggcacc
     1681 aggatggtct gcaccagctg ccccatctca cactggctcg ggacacccta catttccttg
     1741 tcctccgctg gggccacatc cgtcttcctg cctccagcta ccaagacaca gccacccagt
     1801 tctcttcctt agacacccag cttttgatga cactgtatgt ggggaaagaa gaagctgggt
     1861 tctatgtctc taaagcactg gtccacgctg gggtggccct cgtgccccgt ggactgaccc
     1921 tggcacccat ggacggtccc acaacagatg aggtgacact tcaagtctcg ggggagcgag
     1981 agggctcacc cagcactgct gtcagatacc cctcaggcag cgtggccctc cccagccagt
     2041 ggctactcat tggataccac gaacctcccc ccgttctaca cacaaccatg ctgcgggttc
     2101 atcccatccc agggaaagtg tctgctgaga cacgagcctc agaggacctc catgctccac
     2161 ctgtcttttt tgagctccta aacctgagga gggaggatcc agaactacat cctgaagaga
     2221 aagcttcaga ctcttatcca gggctgggat cccaagacct gctagcagct accttcactg
     2281 ccatcctcct gggagcctgg gtcctctacc tgatgaggca gcagcagcag agcccttcag
     2341 cacctgctgg ccctcctgac ctctcacagg atgctcaggg acagctctca agggacatct
     2401 tacaggacca gaggagattt caaagcccct cagagccagc ccagccacct catgaccctg
     2461 aagggcagcc caccgtggtg gggaagattt ccttcaaccc caaggatgtg ctgggccgtg
     2521 gggcaggagg gacttttgtt ttccgaggac agtttgaggg gcgggcagta gctgtggcca
     2581 ggctcctccg agagtgcttt ggcttggtac ggcgagaggt gcagctgctg caagagtctg
     2641 accggcaccc caacgtgctt cgctacttct gcactgagca tggtccccag ttccactaca
     2701 ttgccttgga gctctgccag gcctccttgc aagagtacgt ggagagccca gacctagatc
     2761 gctggggcct ggagcccacc acagtgctgc agcagatgat gtccggcctg gcccatctac
     2821 atttcttaca catagtacac cgtgacctaa agccagccaa cattctcatg gctgggcctg
     2881 acagccaggg ccaaggcaga gtggtcatct ctgacttcgg cctctgcaag aagctacctg
     2941 ttggccgttg tagcttcagt ctccattctg gtattcccgg cacagaaggc tggatggcac
     3001 tagagctcct gcagctcccc ccagacagcc cgaccagcgc tgtggatatc ttctccgcag
     3061 gctgcgtatt ttattacgtg ctttccggtg gcagccaccc ctttggagag agtctctacc
     3121 gacaggccaa catcctctca ggggatccct gtctggctca gctgcaggaa gagacccatg
     3181 acaaggttgt ggcattggac ttagtgagag ccatgctgag cttgctgccc caggatcgcc
     3241 cctcggcagg atgggtactg gctcaccccc tcttttggag cagagctaag gagctccagt
     3301 tcttccagga tgtcagtgac tggttggaga aggaaccaga tcaggggcct ctcgtgtcgg
     3361 cgctggaggc aggaagctac aaggtggtcc gggaagactg gcacaagcac atctcagccc
     3421 ctctgcaggc agatctgaaa aggttccgct catacaaagg gacatcagtt cgagacctgc
     3481 tccgggccat gaggaacaag aagcaccact acagggagct cccagctgag gtgcgccaga
     3541 cactaggcca acttcctgct ggcttcattc agtacttcac acagcgattc cccaggctgt
     3601 tgctgcacac tcaccgtgcc atgaggacct gcgcctctga gagcctcttc ctgccctact
     3661 acccaccagc tctggaagcc aggaggccag atgccacaaa gagcggaggc gagcagaagc
     3721 tgatctctga ggaggatctg tgactcgagc atgcatctag agggccctat tctatagtgt
     3781 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3841 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3901 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3961 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4021 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4081 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4141 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4381 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4441 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4501 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4561 ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4621 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4681 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4741 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4801 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4861 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4921 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5041 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5101 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5221 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5341 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5461 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5701 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5761 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5821 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5881 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5941 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6001 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6061 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6121 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6181 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6901 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
