LOCUS       sequence-316904-        5253 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 5253)
  AUTHORS   Wu R, Shen Q, Newburger PE
  TITLE     Recognition and binding of the human selenocysteine insertion 
            sequence by nucleolin.
  JOURNAL   J Cell Biochem. 2000 Apr . 77(3):507-16.
  PUBMED    10760958
REFERENCE   2  (bases 1 to 5253)
  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: "J Cell 
            Biochem. 2000 Apr . 77(3):507-16."
COMMENT     Sequence Label: pRSET Nucleolin
FEATURES             Location/Qualifiers
     source          1..5253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     467..486
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        467..485
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             517..539
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             559..576
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    1153..1164
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    1171..1182
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             2230..2241
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3101..3119)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3115..3162
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3259..3714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3346..3365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3556..3577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3740..3844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3845..3913,3914..4705)
                     /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(4063..4082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4876..5253,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcaggat ctcgatcccg cgaaattaat acgactcact
      481 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
      541 atacatatgc ggggttctca tcatcatcat catcatggta tggctagcat ggtgaagctc
      601 gcgaaggcag gtaaaaatca aggtgacccc aagaaaatgg ctcctcctcc aaaggaggta
      661 gaagaagata gtgaagatga ggaaatgtca gaagatgaag aagatgatag cagtggagaa
      721 gaggtcgtca tacctcagaa gaaaggcaag aaggctgctg caacctcagc aaagaaggtg
      781 gtcgtttccc caacaaaaaa ggttgcagtt gccacaccag ccaagaaagc agctgtcact
      841 ccaggcaaaa aggcagcagc aacacctgcc aagaagacag ttacaccagc caaagcagtt
      901 accacacctg gcaagaaggg agccacacca ggcaaagcat tggtagcaac tcctggtaag
      961 aagggtgctg ccatcccagc caagggggca aagaatggca agaatgccaa gaaggaagac
     1021 agtgatgaag aggaggatga tgacagtgag gaggatgagg aggatgacga ggacgaggat
     1081 gaggatgaag atgaaattga accagcagcg atgaaagcag cagctgctgc ccctgcctca
     1141 gaggatgagg acgatgagga tgacgaagat gatgaggatg acgatgacga tgaggaagat
     1201 gactctgaag aagaagctat ggagactaca ccagccaaag gaaagaaagc tgcaaaagtt
     1261 gttcctgtga aagccaagaa cgtggctgag gatgaagatg aagaagagga tgatgaggac
     1321 gaggatgacg acgacgacga agatgatgaa gatgatgatg atgaagatga tgaggaggag
     1381 gaagaagagg aggaggaaga gcctgtcaaa gaagcacctg gaaaacgaaa gaaggaaatg
     1441 gccaaacaga aagcagctcc tgaagccaag aaacagaaag tggaaggcac agaaccgact
     1501 acggctttca atctctttgt tggaaaccta aactttaaca aatctgctcc tgaattaaaa
     1561 actggtatca gcgatgtttt tgctaaaaat gatcttgctg ttgtggatgt cagaattggt
     1621 atgactagga aatttggtta tgtggatttt gaatctgctg aagacctgga gaaagcgttg
     1681 gaactcactg gtttgaaagt ctttggcaat gaaattaaac tagagaaacc aaaaggaaaa
     1741 gacagtaaga aagagcgaga tgcgagaaca cttttggcta aaaatctccc ttacaaagtc
     1801 actcaggatg aattgaaaga agtgtttgaa gatgctgcgg agatcagatt agtcagcaag
     1861 gatgggaaaa gtaaagggat tgcttatatt gaatttaaga cagaagctga tgcagagaaa
     1921 acctttgaag aaaagcaggg aacagagatc gatgggcgat ctatttccct gtactatact
     1981 ggagagaaag gtcaaaatca agactataga ggtggaaaga atagcacttg gagtggtgaa
     2041 tcaaaaactc tggttttaag caacctctcc tacagtgcaa cagaagaaac tcttcaggaa
     2101 gtatttgaga aagcaacttt tatcaaagta ccccagaacc aaaatggcaa atctaaaggg
     2161 tatgcattta tagagtttgc ttcattcgaa gacgctaaag aagctttaaa ttcctgtaat
     2221 aaaagggaaa ttgagggcag agcaatcagg ctggagttgc aaggacccag gggatcacct
     2281 aatgccagaa gccagccatc caaaactctg tttgtcaaag gcctgtctga ggataccact
     2341 gaagagacat taaaggagtc atttgacggc tccgttcggg caaggatagt tactgaccgg
     2401 gaaactgggt cctccaaagg gtttggtttt gtagacttca acagtgagga ggatgccaaa
     2461 gctgccaagg aggccatgga agacggtgaa attgatggaa ataaagttac cttggactgg
     2521 gccaaaccta agggtgaagg tggcttcggg ggtcgtggtg gaggcagagg cggctttgga
     2581 ggacgaggtg gtggtagagg aggccgagga ggatttggtg gcagaggccg gggaggcttt
     2641 ggagggcgag gaggcttccg aggaggcaga ggaggaggag gtgaccacaa gccacaagga
     2701 aagaagacga agtttgaata gcttctgtcc ctctgctttc ccttttccat ttgaaagaaa
     2761 ggactctggg gtttttactg ttacctgatc aatgacagag ccttctgagg acattccaag
     2821 acagtataca gtcctgtggt ctccttggaa atccgtctag ttaacatttc aagggcaata
     2881 ccgtgttggt tttgactgga tattcatata aactttttaa agagttgagt gatagagcta
     2941 acccttatct gtaagttttg aatttatatt gtttcatccc atgtacaaaa ccattttttc
     3001 ctacggatcc gagctcgaga tctgcagctg gtaccatgga attcgaagct tgatccggct
     3061 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     3121 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     3181 tccggatctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3241 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3301 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3361 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3421 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3481 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3541 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3601 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3661 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3721 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3841 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3901 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3961 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4021 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4081 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4141 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4201 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4261 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4321 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4381 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4441 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4501 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4561 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4621 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4681 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4741 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5221 cttggagcga acgacctaca ccgaactgag ata
//
