LOCUS       sequence-342084-        6330 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 6330)
  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 6330)
  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: XBP-1 2: pFLAG.XBP1p.CMV2
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2671..3293
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2870..2889)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3317..3337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3322..3651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3502..3637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3689..3708)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3690..3708)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3722..3738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3722..3738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3735..3757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3746..3762
                     /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(3770..3776,3777..3794,3795..3800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3815..3836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3953..3970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4124..4712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4204..4223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4883..5674,5675..5743))
                     /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     5506..5525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5744..5848)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5875..6303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5962..5981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6172..6193
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgtggtggca gcggcgccga gcgcggccac ggcggccccc aaagtgctac tcttatctgg
     1021 ccagcccgcc tccgtcggcc gggcgctgcc gctcatggta cccggtccgc gggcagcagg
     1081 gtcggaggcg agcgggacac cgcaggctcg caagcggcag cggctcacgc acctgagccc
     1141 ggaggagaaa gcgctgcgga ggaaactgaa aaacagagta gtagcgcaga ctgctcgaga
     1201 tagaaagaaa gcccggatga gcgagctgga gcagcaagtg gtggatttgg aagaagagaa
     1261 ccacaaactc cagctagaaa atcagctttt acgggagaaa actcacggcc ttgtggttga
     1321 gaaccaggag ttaagaacac gcttgggaat ggacacgctg gatcctgacg aggttccaga
     1381 ggtggaggcc aaggggagtg gagtaaggct ggtggccggg tctgctgagt ccgcagcagg
     1441 tgcaggccca gttgtcacct ccccagaaca tcttcccatg gactctgaca ctgttgcctc
     1501 ttcagattct gagtctgata tccttttggg cattctggac aagttggacc ctgtcatgtt
     1561 tttcaaatgt ccttccccag agtctgctag tctggaggaa ctcccagagg tctacccaga
     1621 aggacctagt tccttaccag cctccctttc tctgtcagtg gggacctcat cagccaagct
     1681 ggaagccatt aatgaactca ttcgttttga ccatgtatac accaagcctc tagttttaga
     1741 gatcccctct gagacagaga gtcaaactaa cgtggtagtg aaaattgagg aagcacctct
     1801 aagctcttca gaagaggatc accctgaatt cattgtctca gtgaagaaag agcctttgga
     1861 agatgacttc atcccagagc tgggcatctc aaacctgctt tcatccagcc attgtctgag
     1921 accaccttct tgcctgctgg acgctcacag tgactgtgga tatgagggct ccccttctcc
     1981 cttcagtgac atgtcttctc cacttggtac agaccactcc tgggaggata cttttgccaa
     2041 tgaacttttc ccccagctga ttagtgtcta aagagccaca taacactggg cccctttccc
     2101 tgaccatcac attgcctaga ggatagcata ggcctgtctc tttcgttaaa agccaaagta
     2161 gaggctgtct ggccttagaa gaattcctct aaagtatttc aaatctcata gatgacttcc
     2221 aagtattgtc gtttgacact cagctgtcta aggtattcaa aggtattcca gtactacagc
     2281 ttttgagatt ctagtttatc ttaaaggtgg tagtatactc taaatcgcag ggagggtcat
     2341 ttgacagttt tttcccagcc tggcttcaaa ctatgtagcc gaggctaggc agaaacttct
     2401 gaccctcttg accccacctc ccaagtgctg ggcttcacca ggtgtgcacc tccacacctg
     2461 ccccccgaca tgtcaggtgg acatgggatt catgaatggc ccttagcatt tctttctcca
     2521 ctctctgctt cccaggtttc gtaacctgag ggggcttgtt ttcccttatg tgcattttaa
     2581 atgaagatca agaatctttg taaaatgatg aaaatttact atgtaaatgc ttgatggatc
     2641 ttcttgctag tgtagcttct agaggatccc gggtggcatc cctgtgaccc ctccccagtg
     2701 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     2761 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     2821 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     2881 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     2941 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3001 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3061 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3121 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     3181 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg
     3241 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg
     3301 cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3421 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3481 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3541 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3601 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc
     3661 gaggaactga aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca
     3721 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3781 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3841 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3961 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4021 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4081 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4561 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4861 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4921 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4981 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5041 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5101 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5161 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5221 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5281 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5341 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5401 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5461 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5521 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5581 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5641 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5701 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5761 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5821 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     5881 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5941 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6001 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6061 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6121 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6181 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6241 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6301 attttaacaa aatattaacg cttacaattt
//
