LOCUS       sequence-10025-e        6352 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 6352)
  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 6352)
  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 1: pFLAG.XBP1u.CMV2
FEATURES             Location/Qualifiers
     source          1..6352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..150
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     142..159
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     143..159
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        312..690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        691..894
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     845..865
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             928..951
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2691..3315
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3337..3357)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3342..3672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3523..3658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3710..3729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3711..3729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3743..3759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3743..3759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3756..3778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3767..3783
                     /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(3791..3797,3798..3815,3816..3821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3836..3857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3974..3991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4904..5695,5696..5764))
                     /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     5527..5546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5765..5869)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5896..6352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5983..6002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6193..6214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccca
      121 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat ctaatcaata
      181 ttggccatta gccatattat tcattggtta tatagcataa atcaatattg gctattggcc
      241 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
      301 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      361 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      421 cgaccgccca gcgacccccg cccgttgacg tcaatagtga cgtatgttcc catagtaacg
      481 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      541 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      601 tggcccgcct agcattatgc ccagtacatg accttacggg agtttcctac ttggcagtac
      661 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      721 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      781 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg
      841 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      901 gtgaaccgtc agaattgatc taccatggac tacaaagacg atgacgacaa gcttgtggtg
      961 gcagcggcgc cgagcgcggc cacggcggcc cccaaagtgc tactcttatc tggccagccc
     1021 gcctccggcg gccgggcgct gccgctcatg gtacccggtc cgcgggcagc agggtcggag
     1081 gcgagcggga caccgcaggc tcgcaagcgg cagcggctca cgcacctgag cccggaggag
     1141 aaagcgctgc ggaggaaact gaaaaacaga gtagcagcgc agactgctcg agatagaaag
     1201 aaagcccgga tgagcgagct ggagcagcaa gtggtggatt tggaagaaga gaaccacaaa
     1261 ctccagctag aaaatcagct tttacgggag aaaactcacg gccttgtggt tgagaaccag
     1321 gagttaagaa cacgcttggg aatggacacg ctggatcctg acgaggttcc agaggtggag
     1381 gccaagggga gtggagtaag gctggtggcc gggtctgctg agtccgcagc actcagacta
     1441 tgtgcacctc tgcagcaggt gcaggcccag ttgtcacctc cccagaacat cttcccatgg
     1501 actctgacac tgttgcctct tcagattctg agtctgatat ccttttgggc attctggaca
     1561 agttggaccc tgtcatgttt ttcaaatgtc cttccccaga gtctgctagt ctggaggaac
     1621 tcccagaggt ctacccagaa ggacctagtt ccttaccagc ctccctttct ctgtcagtgg
     1681 ggacctcatc agccaagctg gaagccatta atgaactcat tcgttttgac catgtataca
     1741 ccaagcctct agttttagag atcccctctg agacagagag tcaaactaac gtggtagtga
     1801 aaattgagga agcacctcta agctcttcag aagaggatca ccctgaattc attgtctcag
     1861 tgaagaaaga gcctttggaa gatgacttca tcccagagct gggcatctca aacctgcttt
     1921 catccagcca ttgtctgaga ccaccttctt gcctgctgga cgctcacagt gactgtggat
     1981 atgagggctc cccttctccc ttcagtgaca tgtcttctcc acttggtaca gaccactcct
     2041 gggaggatac ttttgccaat gaacttttcc cccagctgat tagtgtctaa agagccacat
     2101 aacactgggc ccctttccct gaccatcaca ttgcctagag gatagcatag gcctgtctct
     2161 ttcgttaaaa gccaaagtag aggctgtctg gccttagaag aattcctcta aagtatttca
     2221 aatctcatag atgacttcca agtattgtcg tttgacactc agctgtctaa ggtattcaaa
     2281 ggtattccag tactacagct tttgagattc tagtttatct taaaggtggt agtatactct
     2341 aaatcgcagg gagggtcatt tgacagtttt ttcccagcct ggcttcaaac tatgtagccg
     2401 aggctaggca gaaacttctg accctcttga ccccacctcc caagtgctgg gcttcaccag
     2461 gtgtgcacct ccacacctgc ccccccgaca tgtcaggtgg acatgggatt catgaatggc
     2521 ccttagcatt tctttctcca ctctctgctt cccaggtttc gtaacctgag ggggcttgtt
     2581 ttcccttatg tgcattttaa atgaagatca agaatctttg taaaatgatg aaaatttact
     2641 atgtaaatgc ttgatggatc ttcttgctag tgtagcttct agaggatccc gggtggcatc
     2701 cctgtgaccc ctccccagtg cctctcctgg ccttggaagt tgccactcca gtgcccacca
     2761 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc tctataatat
     2821 tatggggtgg aggggggtgg tatggagcaa ggggcccaag ttgggaagac aacctgtagg
     2881 gcctgcgggg tctattcggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     2941 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     3001 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3061 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     3121 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa
     3181 ataactatac cagcaggagg acgtccagac acagcatagg ctacctgcca tggcccaacc
     3241 ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta
     3301 gatgcctgtt gaattgggta cgcggccagc ttctgtggaa tgtgtgtcag ttagggtgtg
     3361 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3421 caaccaggtg tggaaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3481 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3541 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3601 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3661 ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga
     3721 gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3781 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3841 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3901 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3961 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4981 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5041 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5101 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5161 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5221 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5281 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5341 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5401 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5461 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5521 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5581 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5641 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5701 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5761 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5821 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5881 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5941 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6001 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     6061 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6121 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6181 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6241 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6301 tttaacaaaa atttaacgcg aattttaaca aaatattaaa cgtttacaat tt
//
