LOCUS       sequence-224395-        6989 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 6989)
  AUTHORS   Yang Y, Fang S, Jensen JP, Weissman AM, Ashwell JD
  TITLE     Ubiquitin protein ligase activity of IAPs and their degradation in 
            proteasomes in response to apoptotic stimuli.
  JOURNAL   Science 2000 May 5;288(5467):874-7.
  PUBMED    10797013
REFERENCE   2  (bases 1 to 6989)
  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: "Science"; 
            date: "2000-05-5"; volume: "288(5467)"; pages: "874-7"
COMMENT     Sequence Label: pEBB-XIAP
FEATURES             Location/Qualifiers
     source          1..6989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        201..411
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        223..419
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        506..1684
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          737..1675
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1632..1652
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1740..1763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4066..4083)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4066..4082)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4075..4097)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4295..4750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4382..4401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4592..4613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4767..4786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4886..4908
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4946..4964)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5032..5136
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5137..5205,5206..5997)
                     /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(5355..5374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6168..6756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6657..6676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6910..6927
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      181 caagcttaat tccctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt
      481 ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc
      541 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
      601 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
      661 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
      721 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
      781 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
      841 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      901 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      961 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     1021 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     1081 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     1141 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     1201 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     1261 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     1321 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     1381 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1441 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1501 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1561 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1621 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1681 gtgaggaatt ctctagagat ccctcgacct cgagatccat tgtgctggat ctcaccatgg
     1741 actacaaaga cgatgacgac aagggatcca tgacttttaa cagttttgaa ggatctaaaa
     1801 cttgtgtacc tgcagacatc aataaggaag aagaatttgt agaagagttt aatagattaa
     1861 aaacttttgc taattttcca agtggtagtc ctgtttcagc atcaacactg gcacgagcag
     1921 ggtttcttta tactggtgaa ggagataccg tgcggtgctt tagttgtcat gcagctgtag
     1981 atagatggca atatggagac tcagcagttg gaagacacag gaaagtatcc ccaaattgca
     2041 gatttatcaa cggcttttat cttgaaaata gtgccacgca gtctacaaat tctggtatcc
     2101 agaatggtca gtacaaagtt gaaaactatc tgggaagcag agatcatttt gccttagaca
     2161 ggccatctga gacacatgca gactatcttt tgagaactgg gcaggttgta gatatatcag
     2221 acaccatata cccgaggaac cctgccatgt atagtgaaga agctagatta aagtcctttc
     2281 agaactggcc agactatgct cacctaaccc caagagagtt agcaagtgct ggactctact
     2341 acacaggtat tggtgaccaa gtgcagtgct tttgttgtgg tggaaaactg aaaaattggg
     2401 aaccttgtga tcgtgcctgg tcagaacaca ggcgacactt tcctaattgc ttctttgttt
     2461 tgggccggaa tcttaatatt cgaagtgaat ctgatgctgt gagttctgat aggaatttcc
     2521 caaattcaac aaatcttcca agaaatccat ccatggcaga ttatgaagca cggatcttta
     2581 cttttgggac atggatatac tcagttaaca aggagcagct tgcaagagct ggattttatg
     2641 ctttaggtga aggtgataaa gtaaagtgct ttcactgtgg aggagggcta actgattgga
     2701 agcccagtga agacccttgg gaacaacatg ctaaatggta tccagggtgc aaatatctgt
     2761 tagaacagaa gggacaagaa tatataaaca atattcattt aactcattca cttgaggagt
     2821 gtctggtaag aactactgag aaaacaccat cactaactag aagaattgat gataccatct
     2881 tccaaaatcc tatggtacaa gaagctatac gaatggggtt cagtttcaag gacattaaga
     2941 aaataatgga ggaaaaaatt cagatatctg ggagcaacta taaatcactt gaggttctgg
     3001 ttgcagatct agtgaatgct cagaaagaca gtatgccaga tgagtcaagt cagacttcat
     3061 tacagaaaga gattagtact gaagagcagc taaggcgcct gcaagaggag aagctttgca
     3121 aaatctgtat ggatagaaat attgctatcg tttttgttcc ttgtggacat ctagtcactt
     3181 gtaaacaatg tgctgaagca gttgacaagt gtcccatgtg ctacacagtc attactttca
     3241 agcaaaaaat ttttatgtct taatctaact ctatagtagg catgttatgt tgttcttatt
     3301 accctgattg aatgtgtgat gtgaactgac tttaagtaat caggattgaa ttcgatatca
     3361 agcttatcga ttgactgact gagcggccgc gactctagag tgagggtccc cacctgggac
     3421 ccttgagagt atcaggtctc ccacgtggga gacaagaaat ccctgtttaa tatttaaaca
     3481 gcagtgttcc ccatctgggt ccttgcaccc ctcactctgg cctcagccga ctgcacagcg
     3541 gcccctgcat ccccttggct gtgaggcccc tggacaagca gaggtggcca gagctgggag
     3601 gcatggccct ggggtcccac gaatttgctg gggaatctcg tttttcttct taagactttt
     3661 gggacatggt ttgactcccg aacatcaccg acgtgtctcc tgtttttctg ggtggcctcg
     3721 ggacacctgc cctgccccca cgagggtcag gactgtgact ctttttaggg ccaggcaggt
     3781 gcctggacat ttgccttgct ggacggggac tggggatgtg ggagggagca gacaggagga
     3841 atcatgtcag gcctgtgtgt gaaaggaagc tccactgtca ccctccacct cttcaccccc
     3901 cactcaccag tgtcccctcc actgtcacat tgtaactgaa cttcaggata ataaagtgtt
     3961 tgcctccaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4021 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaccg aattcactgg ccgtcgtttt
     4081 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4141 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4201 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4261 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4321 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4381 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4441 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4501 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4561 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4621 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4681 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4741 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4801 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4861 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4921 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4981 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5041 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5101 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5161 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5221 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5281 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5341 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5401 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5461 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5521 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5581 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5641 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5701 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5761 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5821 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5881 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5941 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6001 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6061 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6121 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6181 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6241 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6301 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6361 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6421 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6481 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6541 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6601 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6661 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6721 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6781 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6841 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6901 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6961 ctccccgcgc gttggccgat tcattaatg
//
