LOCUS       sequence-255005-        6308 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 6308)
  AUTHORS   Boothroyd CE, Dreesen O, Leonova T, Ly KI, Figueiredo LM, Cross GA, 
            Papavasiliou FN
  TITLE     A yeast-endonuclease-generated DNA break induces antigenic switching
            in Trypanosoma brucei.
  JOURNAL   Nature. 2009 May 14;459(7244):278-81. doi: 10.1038/nature07982. Epub
            2009 Apr 15.
  PUBMED    19369939
REFERENCE   2  (bases 1 to 6308)
  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"; doi: 
            "10.1038/nature07982"; journalName: "Nature"; date: "2009-05-14- 
            14"; volume: "459"; issue: "7244"; pages: "278-81"
COMMENT     Sequence Label: pLew100::NLS-ISceI-HA
FEATURES             Location/Qualifiers
     source          1..6308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    19..37
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             167..187
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             896..922
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1657..1675)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1658..1675)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1965..1984)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2194..2213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2313..2335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2373..2391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2459..2563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2564..2632,2633..3424)
                     /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(2782..2801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3595..4183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4084..4103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4337..4354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(5510..5884)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(6023..6042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6024..6042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6306..6308,1..16)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ctatcagtga tagagatctc cctatcagtg atagagatcc ctgagtactg agtttaacat
       61 gttctcgtcc cgggctgcac gcgccttcga gttttttttc cttttcccca tttttttcaa
      121 cttgaagact tcaattacac caaaaagtaa aattcacaag cttatgccaa agaagaagcg
      181 aaaggtacat atgaaaaaca tcaaaaaaaa ccaggtaatg aacctgggtc cgaactctaa
      241 actgctgaaa gaatacaaat cccagctgat cgaactgaac atcgaacagt tcgaagcagg
      301 tatcggtctg atcctgggtg atgcttacat ccgttctcgt gatgaaggta aaacctactg
      361 tatgcagttc gagtggaaaa acaaagcata catggaccac gtatgtctgc tgtacgatca
      421 gtgggtactg tccccgccgc acaaaaaaga acgtgttaac cacctgggta acctggtaat
      481 cacctggggc gcccagactt tcaaacacca agcattcaac aaactggcta acctgttcat
      541 cgttaacaac aaaaaaacca tcccgaacaa cctggttgaa aactacctga ccccgatgtc
      601 tctggcatac tggttcatgg atgatggtgg taaatgggat tacaacaaaa actctaccaa
      661 caaatcgatc gtactgaaca cccagtcttt cactttcgaa gaagtagaat acctggttaa
      721 gggtctgcgt aacaaattcc aactgaactg ttacgtaaaa atcaacaaaa acaaaccgat
      781 catctacatc gattctatgt cttacctgat cttctacaac ctgatcaaac cgtacctgat
      841 cccgcagatg atgtacaaac tgccgaacac tatctcctcc gaaactttcc tgaaataccc
      901 atacgatgtt ccagattacg cttgatcctg cccatttagt tggcttttcc cttgtctcgt
      961 gtcttttccg tggaaaggtt cccggagtaa tctgatggca cagcagggag gtgcgcctgc
     1021 aggttggtta ggaagggggg atgatgtaaa agaagaaaat ggggggatat cttatgttta
     1081 agaggataaa taatgtgaag gggctttata tgcttgcttg gttgtctcgt tggtgcatgg
     1141 ggaatctgca tgtttgcttt ggagcacgcg tggtacatca atgggatcat atcttgctgc
     1201 ctcccgcagt caccgtgcgg agctgccggt gccccgtttc tcataggcgt gtgcaccttc
     1261 cgtctgagac ctgtaaatgg ttaaaaagga atatataatt accttttgaa aagtggtaaa
     1321 aaacgaatat atgttttttt ttggttttaa tacgctcttt tgtgtgtatg agaggaataa
     1381 ataagtgtgt gtgtgtgtgt gtgttgttgt tgttgaaaaa atgaaggaca cgaaagcgct
     1441 cattaagggt ggggatacac caggcataat tatccgaaaa tattttgatg caccaaataa
     1501 gtgaacatac tgacgaaatc aaagagttgg aggatggata gggaggcctc attggcagtg
     1561 gtaaattgat ttgacttgaa tatgataccc tcttactgtt tcattgccac tacaagttgg
     1621 tttccttccc ctgcaggcat gcaagctagc ttgtattcta tagtgtcacc taaatcgtat
     1681 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaca atatgtacaa
     1741 gcctaattgt gtagcatctg gcttactgaa gcagacccta tcatctctct cgtaaactgc
     1801 cgtcagagtc ggtttggttg gacgaacctt ctgagtttct ggtaacgccg tcccgcaccc
     1861 ggaaatggtc agcgaaccaa tcagcagggt catcgctagc cagatcctct acgccggacg
     1921 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     1981 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     2041 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc accattcctt
     2101 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     2161 cataagggag agcgtcgaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2221 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2281 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2341 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     2401 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2461 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2521 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2581 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2641 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2701 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2761 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2821 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2881 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2941 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3001 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3061 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3121 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3181 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3241 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3301 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3361 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3421 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3481 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3541 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3601 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3661 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3721 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3781 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3841 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3901 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3961 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4021 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4081 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4141 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4201 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4261 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4321 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4381 accgcctctc cccgcgcgtt ggccgattca ttaatgcaga attcgagctc atatagttgg
     4441 tatgtattct aattccagac tactggcgtg ggataaacat gtccccctga ttaaaaggaa
     4501 agattccata gcccattagt gcaaagataa ttggtacacc gtcaaaaaca catgggagac
     4561 cacacgatac gacactgtga ccgtagcatc aattccgcac tcgatataac tctatcgaag
     4621 aacttccatg gtacaaactg gtcgccacgg tctgctccac acggagatca tcgtatcatt
     4681 tttatcgata gcggccgcta tcgatgtatg ccttggccct gatggcatgc caatttcact
     4741 acaacggact atgtggaccc cgttatcatg gaaatgcgct agttggagga agttagaccg
     4801 cgccggaaaa gagaggggta gagaaaatga caacttggaa gatatccaca cacgcacggt
     4861 gaaacgttag caacaattat tagggaagcg acgcttgccc tagtcccacg agtaaccaag
     4921 actccaaaag cctttctggc acagagagcg agccgaaatg gaaaagagaa acaatgcctg
     4981 cactaacact actgagcgat tcgcctcgcc gcggaggacc gaatactaat aacgacactt
     5041 gcggtcaaaa agtagaagaa caaatgctca acgatgagtg aatcaggtta gggtagttgg
     5101 aaaattatac agaatgtctt tggcaacaca ccgggcttgc atgcctgcaa ggccttgcaa
     5161 ggccttgcag aatactgcat agataacaaa cgcatcaaac acaactggtg gcactatttc
     5221 aatcatgtcg acacaccaag acgagaaaat ttgagttaaa caagatgcct acaaccctat
     5281 atttattatc ttcaagtgat cttctcttct cttgtgcgct gtacgtaaat gtgttgcaaa
     5341 taaggcatag ggctgagtac aggcaccaca cgcgaggaaa atcataagca ctagccgtgt
     5401 caactcacag cgactacaga acaattttgg ccacacaacc cggtgttagg atctccgagg
     5461 cctcggaccc gtcgggccgc gtcggaccgg cggtgttggt cggcgtcggt cagtcctgct
     5521 cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc
     5581 acggctgctc gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca
     5641 cgacctccga ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg
     5701 tgttgtccgg caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga
     5761 ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga
     5821 actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg
     5881 ccatggtgat agcttatttt atggcagcaa cgagacctta cgtaaagcaa aactaaataa
     5941 gagcggagac tgcaatgcag agtaaaaaaa aaaaaaacat gaatttcaga agaccttgct
     6001 gtgcccccgg tacgggagat ctccctatag tgagtcgtat taatcaggta ccgtcattgg
     6061 ggttaagcgg aaaggtgtgt gtcagtaggt tgtgaggtga aagcgttttc agatgcatag
     6121 tgagcttaat gtccttttca cagtatatcg tgtctgatag gtatctctta ttagtatagt
     6181 cgaatactag tcaatagtgc gttttgtgca aaatgtccat tttgtggcag tgatggggtt
     6241 gttttatgct attccgtgtc tctgggtggg cgtgcattga aaataggggt tatcgggtag
     6301 ggatctcc
//
