LOCUS       sequence-10070-e        5601 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 5601)
  AUTHORS   Calfon M, Zeng H, Urano F, Till JH, Hubbard SR, Harding HP, Clark 
            SG, Ron D
  TITLE     IRE1 couples endoplasmic reticulum load to secretory capacity by 
            processing the XBP-1 mRNA.
  JOURNAL   Nature. 2002 Jan 3. 415(6867):92-6.
  PUBMED    11780124
REFERENCE   2  (bases 1 to 5601)
  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"; journalName: "Nature. 
            2002 Jan 3. 415(6867):92-6."
COMMENT     Sequence Label: BiP 2: C. elegans BiP (hsp-4)-GFP
FEATURES             Location/Qualifiers
     source          1..5601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1271..1299)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2047..2067
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3041..3063)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3163..3182
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3260..3278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3346..3450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3451..3519,3520..4311)
                     /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(3669..3688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4482..5070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4971..4990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5224..5241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5358..5379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5394..5399,5400..5417,5418..5424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5432..5448
                     /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)."
     primer_bind     5437..5459
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5456..5472
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5456..5472
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgacccgttt gactactcca aattcacgaa gttaccaaaa
       61 taaaagtgat atttgacttt cagaaatacc atttcaaatt ccctaagacg ctcgggaaat
      121 attaattact gcaatttata ttctgcttgt atttttcgaa gttgggtcca actgtgtgaa
      181 gtattgtaag aatcatatcc ttctccttca cattctacat aaacaattca tttctattct
      241 gtaaattttt tctgatgatt tacggtaaaa acgagcgaaa ttcggtccgg gacaagggtt
      301 tctacgacga gtccatcgtg gtgccgctcg cttgtttgaa ttcccgcgtg ccgcattcct
      361 cgtgtcgaga cccgatgtcc aactgggggg attaccaact cgggggattg gccccgccca
      421 cagaaccgtg gcttgcaatt ttttcttgtt aattctcgct ctattgagaa aaaataattt
      481 taaaaccgtg cggcagtttc aaaaatgggc gtattgcaag ccacggttct gtgggcgggg
      541 ccaatccccc gagttcttcg ggtctctaag gaaaggattc gtacattctg gtccttttta
      601 tttattttta acctcttttt atttttttaa accgcaatcc attaccagtt ccattttctc
      661 cgtactcgtc agtgtagcga gtgacgagtg aaattgaccc catttcttat cttatcgaaa
      721 acaatctaaa tagtttccgc attcgcataa ccagaaaatt cttcggtagt cgttctcatt
      781 tgtttttatt tcatgaacat aaagtaacgc catagtcttt ttatgaaacg tggcgttaag
      841 aaagctctcg aaaagtctcg atttctccag cactaataca cgtcatctcc gataagtaca
      901 cgttgcatag gcggtcctaa taaaagcgac cgcggacgtt cacattcagt tctttgtttt
      961 ctttttgtcg tctgcactcc ttctttgcga cgtgtatttt gtgttcttct ctgtgtttcc
     1021 acttctcgtt agtattctcg cgcttctact ctgaaaggtt tttcttctta aatgttctca
     1081 tttttcagcc actcagcgaa cagttgaact gaccgctcat caagagaaaa atatgaaaga
     1141 tccccgggat tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga
     1201 acattttcag gaggaccctt ggagggtacc ggtagaaaaa atgagtaaag gagaagaact
     1261 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     1321 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
     1381 ttgcactact ggaaaactac ctgttccatg ggtaagttta aacatatata tactaactaa
     1441 ccctgattat ttaaattttc agccaacact tgtcactact ttctgttatg gtgttcaatg
     1501 cttctcgaga tacccagatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     1561 aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtaa
     1621 gtttaaacag ttcggtacta actaaccata catatttaaa ttttcaggtg ctgaagtcaa
     1681 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
     1741 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat
     1801 ggcagacaaa caaaagaatg gaatcaaagt tgtaagttta aacatgattt tactaactaa
     1861 ctaatctgat ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt
     1921 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     1981 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga
     2041 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     2101 atacaaatag cattcgtaga attccaactg agcgccggtc gctaccatta ccaacttgtc
     2161 tggtgtcaaa aataataggg gccgctgtca tcagagtaag tttaaactga gttctactaa
     2221 ctaacgagta atatttaaat tttcagcatc tcgcgcccgt gcctctgact tctaagtcca
     2281 attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt
     2341 attatcaaaa aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta
     2401 acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat
     2461 tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta
     2521 cacttcttat gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg
     2581 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     2641 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     2701 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     2761 ggggtttccc ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac
     2821 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     2881 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     2941 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     3001 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3061 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3121 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     3181 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3241 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3301 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3481 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3541 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3601 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3661 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3721 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3781 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3841 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3901 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3961 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4021 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4081 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4141 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4201 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4261 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4321 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4381 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4621 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4861 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5101 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5161 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5221 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5281 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5341 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5401 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5461 aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact
     5521 attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg
     5581 ctaacaactt ggaaatgaaa t
//
