LOCUS       sequence-193250-        5714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses UGI in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  AUTHORS   Wang L, Xue W, Yan L, Li X, Wei J, Chen M, Wu J, Yang B, Yang L, 
            Chen J
  TITLE     Enhanced base editing by co-expression of free uracil DNA 
            glycosylase inhibitor.
  JOURNAL   Cell Res. 2017 Aug 29. doi: 10.1038/cr.2017.111.
  PUBMED    28849781
REFERENCE   2  (bases 1 to 5714)
  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: "Cell Res. 
            2017 Aug 29. doi: 10.1038/cr.2017.111."
COMMENT     Sequence Label: pUGI-NLS
FEATURES             Location/Qualifiers
     source          1..5714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..292
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             293..313
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(361..378)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    367..591
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      637..1065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(724..743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     934..955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1074..1094)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1079..1408
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1259..1394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1321..1340
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1627..1646
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1715..2314
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1715..1734)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2211..2231
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2488..2609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2525..2544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2579..2598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2658..2674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2658..2674)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2671..2693)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2682..2698
                     /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(2706..2712,2713..2730,2731..2736))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2751..2772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2889..2906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3060..3648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3140..3159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3819..4610,4611..4679))
                     /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     4442..4461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4680..4784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4859..4878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5050..5429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5430..5633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5584..5604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5678..5697
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5678..5696
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 actcgagcgg ccgccactgt gctggatatc tgcagaattc atgactaatc tgtcagatat
       61 tattgaaaag gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga
      121 ggaggtggaa gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta
      181 cgacgagagc accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc
      241 ttgggctctg gtcatacagg atagcaacgg tgagaacaag attaagatgc tccccaagaa
      301 gaagaggaaa gtctagggat ccgagctcgg taccaagctt aagtttaaac cgctgatcag
      361 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
      421 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
      481 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
      541 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
      601 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
      661 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
      721 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      781 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      841 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      901 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      961 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1021 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     1081 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1141 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     1201 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     1261 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     1321 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     1381 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     1441 atctgatcaa gagacaggat gaggatcgtt tcgcatggaa gcgggtaggc ctttggggca
     1501 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     1561 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     1621 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     1681 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     1741 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     1801 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     1861 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     1921 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     1981 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     2041 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     2101 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     2161 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     2221 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     2281 cacctggtgc atgacccgca agcccggtgc ctgagcggga ctctggggtt cgaaatgacc
     2341 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     2401 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     2461 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     2521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2581 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     2641 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2701 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2761 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2821 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2881 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2941 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3001 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3061 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4801 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     4861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     4921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     4981 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     5041 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     5101 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5161 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     5221 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     5281 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     5341 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5461 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5521 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5581 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     5641 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     5701 cccaagctgg ctag
//
