LOCUS       sequence-10060-e        8314 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 8314)
  TITLE     IRE1 plasmids
REFERENCE   2  (bases 1 to 8314)
  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: IRE1bpcDNA3
FEATURES             Location/Qualifiers
     source          1..8314
                     /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"
     promoter        complement(3866..3884)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3867..3884)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3904..3921)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3910..4134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4180..4608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4267..4286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4477..4498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4617..4637)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4622..4952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4803..4938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4865..4884
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5019..5813
                     /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(5073..5092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5683..5702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5987..6108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6024..6043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6078..6097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6157..6173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6157..6173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6170..6192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6181..6197
                     /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(6205..6211,6212..6229,6230..6235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6250..6271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6388..6405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6559..7147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6639..6658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7318..8109,8110..8178))
                     /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     7941..7960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8179..8283)
                     /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 aagcagggca gcccaccgtg gtggggaaga tttccttcaa ccccaaggat gtgctgggcc
     2521 gtggggcagg agggactttt gttttccgag gacagtttga ggggcgggca gtagctgtga
     2581 agaggctcct ccgagagtgc tttggcttgg ttcggcgaga ggtgcagctg ctgcaagagt
     2641 ctgaccggca ccccaacgtg cttcgctact tctgcactga gcatggtccc cagttccact
     2701 acattgcctt ggagctctgc caggcctcct tgcaagagta cgtggagagc ccagacctag
     2761 atcgctgggg cctggagccc accacagtgc tgcagcagat gatgtccggc ctggcccatc
     2821 tacattcctt acacatagta caccgtgacc taaagccagc caacattctc atggctgggc
     2881 ctgacagcca gggccaaggc agagtggtca tctctgactt cggcctctgc aagaagctac
     2941 ctgttggccg ttgtagcttc agtctccatt ctggtattcc cggcacagaa ggctggatgg
     3001 caccagagct cctgcagctc cccccagaca gcccgaccaa cgctgtggat atcttctccg
     3061 caggctgcgt attttattac gtgctttccg gtggcagcca cccctttgga gagagtctct
     3121 accgacaggc caacatcctc tcaggggatc cctgtctggc tcagctgcag gaagagaccc
     3181 atgacaaggt tgtggcattg gacttagtga gagccatgct gagcttgctg ccccaggatc
     3241 gcccctcggc aggatgggta ctggctcacc ccctcttttg gagcagagct aaggagctcc
     3301 agttcttcca ggatgtcagt gactggttgg agaaggaacc agatcagggg cctctcgtgt
     3361 cggcgctgga ggcaggaagc tacaaggtgg tccgggaaga ctggcacaag cacatctcag
     3421 cccctctgca ggcagatctg aaaaggttcc gctcatacaa agggacatca gttcgagacc
     3481 tgctccgggc catgaggaac aagaagcacc actacaggga gctcccagct gaggtgcgcc
     3541 agacactagg ccaacttcct gctggcttca ttcagtactt cacacagcga ttccccaggc
     3601 tgttgctgca cactcaccgt gccatgagga cctgcgcctc tgagagcctc ttcctgccct
     3661 actacccacc agctctggaa gccaggaggc cagatgccac aaagagctga gacatccaga
     3721 gggtccaaag gaggaccaag gagttgagcc tgtggctcag tctagtctca ggcgtgacag
     3781 aaaagatggg cagcggtcta agggactcag gctgttaatg caaaaaaaaa aaaaaaaact
     3841 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     3901 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3961 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4021 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4081 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4141 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4201 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4261 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4321 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4381 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4441 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4501 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     4561 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4621 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     4681 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4861 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4921 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4981 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5041 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5101 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5161 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5221 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5281 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5341 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5401 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5461 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5521 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5581 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5641 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5701 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5761 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5821 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5881 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5941 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6001 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6061 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6121 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6721 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6961 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8281 gcgcacattt ccccgaaaag tgccacctga cgtc
//
