LOCUS       sequence-10063-e        7017 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 7017)
  TITLE     IRE1 plasmids
REFERENCE   2  (bases 1 to 7017)
  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.del-C.9E10.pCDNA3
FEATURES             Location/Qualifiers
     source          1..7017
                     /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             2537..2566
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2607..2624)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2613..2837
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2883..3311
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2970..2989)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3180..3201
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3320..3340)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3325..3655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3506..3641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3568..3587
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3722..4516
                     /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(3776..3795)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4386..4405
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4690..4811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4727..4746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4781..4800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4860..4876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4860..4876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4873..4895)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4884..4900
                     /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(4908..4914,4915..4932,4933..4938))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4953..4974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5091..5108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5262..5850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5342..5361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6021..6812,6813..6881))
                     /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     6644..6663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6882..6986)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcggc
      961 agcaggagac tttgtggttg cccagactat ggcgagacca gtccagagat ttcaactgtg
     1021 gtctcctctc gggttcctgc tgcagcttgt gacgttgctg gggaaactcg gtccacaggt
     1081 ccagtctgtc aggccagaga gccttctgtt tgtgtctacc tgggacggaa gtctccatgc
     1141 nctgaacaag cagacagggg acttgaagtg gacagtgaag gatgacccca tcatccaggg
     1201 ccccatgtat gtcacagaaa tggcctttct ctctgaccca gctgacggca gcctgtatgt
     1261 cttgggaacc cacaaactac aggggctgat gaaactgcca ttcaccatcc cagagctggt
     1321 tcatgcctct ccctgtcgca gctctgatgg tgtcttctac acaggccgga agcaggatgc
     1381 ctggtttgta gtggaccctg agtcagggga gactcagatg acactgacca cagaaggcct
     1441 ctctactcct caacttttca ttggccgaac acagtacaca gtctccatgc atgacctacg
     1501 gaccccagcc ctgcggtgga acacaaccta tcgccgctac tcagcacccc tcctgaatgg
     1561 ctcacccggg aaatacatga gccacctgac atcctgtggg atgggcctgc ttctcactgt
     1621 ggatccagga agtggaattg tgctgtggac acaggacctg ggagtgcctg tgacagggat
     1681 ttacacatgg caccaggatg gtctgcacca gctgccccat ctcacactgg ctcgggacac
     1741 cctacatttc cttgtcctcc gctggggcca catccgtctt cctgcctcca gctaccaaga
     1801 cacagccacc cagttctctt ccttagacac ccagcttttg atgacactgt atgtggggaa
     1861 agaagaagct gggttctatg tctctaaagc actggtccac gctggggtgg ccctcgtgcc
     1921 ccgtggactg accctggcac ccatggacgg tcccacaaca gatgaggtga cacttcaagt
     1981 ctcgggggag cgagagggct cacccagcac tgctgtcaga tacccctcag gcagcgtggc
     2041 cctccccagc cagtggctac tcattggata ccacgaacct ccccccgttc tacacacaac
     2101 catgctgcgg gttcatccca tcccagggaa agtgtctgct gagacacgag cctcagagga
     2161 cctccatgct ccacctgtct tttttgagct cctaaacctg aggagggagg atccagaact
     2221 acatcctgaa gagaaagctt cagactctta tccagggctg ggatcccaag acctgctagc
     2281 agctaccttc actgccatcc tcctgggagc ctgggtcctc tacctgatga ggcagcagca
     2341 gcagagccct tcagcacctg ctggccctcc tgacctctca caggatgctc agggacagct
     2401 ctcaagggac atcttacagg accagaggag atttcaaagc ccctcagagc cagcccagcc
     2461 acctcatgac cctgaagggc agcccaccgt ggtggggaag atttccttca accccaagga
     2521 tgtgctgggc cgtggggagc agaagctgat ctctgaggag gatctgtgac tcgagtctag
     2581 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2641 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2701 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2761 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2821 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2881 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2941 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3001 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3061 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3121 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3181 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3241 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3301 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3361 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3481 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3541 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3601 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3661 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3721 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3781 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3841 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3901 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3961 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4021 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4081 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4141 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4201 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4261 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4321 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4381 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4441 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4501 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4561 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4621 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4681 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4741 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4801 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4861 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4921 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4981 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5041 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5101 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5161 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5221 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5281 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5341 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5401 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     5461 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5521 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5581 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5641 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5701 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5761 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5881 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6061 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6121 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6181 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6241 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6301 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6361 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6421 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6481 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6541 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6601 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6661 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6721 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6781 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6841 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6901 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6961 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
