LOCUS       sequence-443116-        6952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-SV40 NLS-enOgeuIscB-nucleoplasmin NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6952)
  AUTHORS   Yan H, Tan X, Zou S, Sun Y, Ke A, Tang W
  TITLE     Assessing and engineering the IscB-omegaRNA system for programmed 
            genome editing.
  JOURNAL   Nat Chem Biol. 2024 Jul 8. doi: 10.1038/s41589-024-01669-3.
  PUBMED    38977787
REFERENCE   2  (bases 1 to 6952)
  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: "Nat Chem 
            Biol. 2024 Jul 8. doi: 10.1038/s41589-024-01669-3."
COMMENT     Sequence Label: enOgeuIscB
FEATURES             Location/Qualifiers
     source          1..6952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(249..353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(428..447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        619..998
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        999..1202
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1153..1173
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1247..1266
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1247..1265
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1301..1310
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1316..1336
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2834..2881
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(2927..2944)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2933..3157
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3203..3631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3290..3309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3500..3521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3640..3660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3645..3974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3825..3960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3887..3906
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4041..4835
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4095..4114)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4705..4724
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5009..5130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5046..5065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5100..5119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5179..5195)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5179..5195)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5192..5214)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5203..5219
                     /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(5227..5233,5234..5251,5252..5257))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5272..5293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5410..5427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5581..6169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5661..5680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6340..6952,1..179,180..248))
                     /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"
ORIGIN
        1 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
       61 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      361 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
      421 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      481 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      541 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      601 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      661 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      721 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      781 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      841 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      901 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      961 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1021 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1081 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1141 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1201 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     1261 atagggagac ccaagctggc tagcgtttaa acttaagctt gccaccatgg gatctccaaa
     1321 aaagaaacgg aaggtgggat ctatggccgt ggtctacgtg atctctaagt ccggcaaacc
     1381 actgatgccc accaccagat gcggccacgt gcggatcttg ctcaaggaag gcaaggctag
     1441 agtggtggaa agaaagccct tcaccatcca gctgacctac gagtctgccg aggaaaccca
     1501 gcctctggtg ctgggcattg accctggcag aaccaatatc ggtatgagcg tggtgaccga
     1561 gagcggagag agcgtgttca acgcccaaat cgagacaaga aacaaggacg tgcctaagct
     1621 gatgaaggac agaaagcagt ataggcgggc ccacagaagg ctgaagcggc ggtgcaaaag
     1681 acggcggaga gccaaggctg ccggcaccgc cttcgaggag ggcgaaaagc aaagactgct
     1741 gcctggctgc aagaagccta ttacatgcaa gagcatccgg aacaaggaag ccagattcaa
     1801 caaccgcaag cggcctaagg gatggctgac acctaccgcc aatcacctgc tggtcaccca
     1861 cctgaacgtg gttaagaagg tgcagaagat cctgcccgtg gccaaggtgg tgctggagct
     1921 gaatcggttc agctttatgg ccatgaacaa ccccaaggtt cagagatggc agtaccagcg
     1981 gggccctctg tacggcaaag gcagcgtgga agaggccgtg tccatgcagc aggatggaca
     2041 ctgcctgttt tgtaaacacg gcatcgacca ctaccaccac gtggtcccca gaagaaagaa
     2101 cggcagcgag acactggaaa acagagtggg cctttgtgaa gaacaccata gactggtgca
     2161 cacagataag gagtgggagg ccagactggc cagcaagaag tctggaatga acaagaaata
     2221 ccacgccctg tctgtgctga accagatcat cccctacctg gctgatcagc tggccgatat
     2281 gttccccgga aatttctgcg tgacaagcgg ccgggacacc aagctgtttc gggaagagca
     2341 tggcatccct aaggaccact acctggacgc ctactgcatc gcctgcagcg cactgaccga
     2401 cgccaagaag gtgagcagcc ctaagggcag accatacatg gtgagacagt tcagaaggca
     2461 tgatagacag gcctgtcaca aggccaatct gaaccggagc tactacatgg gcggcaagaa
     2521 ggttgccacc aaccggcaca aagctatgga ccagaaaact gacagcctgg aagagtacag
     2581 agccgctcac agcgccgctg acgtgtctaa actgaccgtg aagcaccctt ctgctcagta
     2641 caaggatatg agcagaatca tgcctggcag catcctggtg agcggcgagg gcaaactgtt
     2701 cacactgagc agatctgagg gaagaaacaa aggccaggtg aactacttcg tgtccaccga
     2761 gggcatcaag tactgggcca gaaagtgcca gtatctgcgg aacaacggcg gactgcagat
     2821 ctacgtggga tccaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa
     2881 gtgaggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2941 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3001 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3061 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3121 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3181 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3541 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3601 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3901 ccgaggccgc ctctgcctct gagctattcc agaagtagta aggaggcttt tttggaggcc
     3961 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4021 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4801 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4861 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4921 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4981 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5161 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5221 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5281 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5341 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6001 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cg
//
