LOCUS       sequence-453982-        7102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector encoding human codon-optimized enhanced-activity OgeuIscB 
            driven by CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7102)
  AUTHORS   Xue N, Hong D, Zhang D, Wang Q, Zhang S, Yang L, Chen X, Li Y, Han 
            H, Hu C, Liu M, Song G, Guan Y, Wang L, Zhu Y, Li D
  TITLE     Engineering IscB to develop highly efficient miniature editing tools
            in mammalian cells and embryos.
  JOURNAL   Mol Cell. 2024 Aug 22;84(16):3128-3140.e4. doi: 
            10.1016/j.molcel.2024.07.007. Epub 2024 Aug 2.
  PUBMED    39096898
REFERENCE   2  (bases 1 to 7102)
  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: 
            "10.1016/j.molcel.2024.07.007"; journalName: "Mol Cell"; date: 
            "2024-08-22- 22"; volume: "84"; issue: "16"; pages: "3128-3140.e4"
COMMENT     Sequence Label: IscB-v3
FEATURES             Location/Qualifiers
     source          1..7102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             932..952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2450..2497
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2504..2584
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(2693..2710)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2699..2923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2969..3397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3056..3075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3266..3287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3406..3426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3411..3740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3591..3726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3653..3672
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3807..4601
                     /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(3861..3880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4471..4490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4775..4896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4812..4831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4866..4885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4945..4961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4945..4961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4958..4980)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4969..4985
                     /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(4993..4999,5000..5017,5018..5023))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5038..5059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5176..5193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5347..5935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5427..5446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6106..6897,6898..6966))
                     /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     6729..6748
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6967..7071)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgggatc tccaaaaaag aaacggaagg tgggatctat
      961 ggccgtggtc tacgtgatct ctaagtccgg caaaccactg atgcccacca ccagatgcgg
     1021 ccacgtgcgg atcttgctca aggaaggcaa ggctagagtg gtggaaagaa agcccttcac
     1081 catccagctg acctacgagt ctgccgagga aacccagcct ctggtgctgg gcattgaccc
     1141 tggcagaacc aatatcggta tgagcgtggt gaccgagagc ggagagagcg tgttcaacgc
     1201 ccaaatcaga acaagaaaca aggacgtgcc taagctgatg aagagaagaa agcagtatag
     1261 gatggcccac agaaggctga agcggcggtg caaaagacgg cggagagcca aggctgccgg
     1321 caccgccttc gaggagggcg aaaagcaaag actgctgcct ggctgcttca agcctattac
     1381 atgcaagagc atccggaaca aggaagccag attcaacaac cgcaagcggc ctagaggatg
     1441 gctgacacct accgccaatc acctgctggt cacccacctg aacgtggtta agaaggtgca
     1501 gaagatcctg cccgtggcca aggtggtgct ggagctgaat cggttcagct ttatggccat
     1561 gaacaacccc aaggttcaga gatggcagta ccagcggggc cctctgtacg gcaaaggcag
     1621 cgtggaagag gccgtgtcca tgcagcagga tggacactgc ctgttttgta aacacggcat
     1681 cgaccactac caccacgtgg tccccagaag aaagaacggc agcgagacac tggaaaacag
     1741 agtgggcctt tgtgaagaac accatagact ggtgcacaca gataaggagt gggaggccaa
     1801 cctggccagc aagaagtctg gaatgaacaa gaaataccac gccctgtctg tgctgaacca
     1861 gatcatcccc tacctggctg atcagctggc cgatatgttc cccggaaatt tctgcgtgac
     1921 aagcggccaa gacacctatc tgtttcggga agagcatggc atccctaagg accactacct
     1981 ggacgcctac tgcatcgcct gcagcgcact gaccgacgcc aagaaggtga gcagccctaa
     2041 gggcagacca tacatggtgc accagttcag aaggcatgat agacaggcct gtcacaaggc
     2101 caatctgaac cggagctact acatgggcgg caagctcgtt gccaccaacc ggcacaaagc
     2161 tatggaccag aaaactgaca gcctggaaga gtacagagcc gctcacagcg ccgctgacgt
     2221 gtctaaactg accgtgaagc acccttctgc tcagtacaag gatatgagca gaatcatgcc
     2281 tggcagcatc ctggtgagcg gcgagggcaa actgttcaca ctgagcagat ctgagggaag
     2341 aaacaaaggc caggtgaact acttcgtgtc caccgagggc atcaagtact gggccagaaa
     2401 gtgccagtat ctgcggaaca acggcggact gcagatctac gtgggatcca aaaggccggc
     2461 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga tcttacccat acgatgttcc
     2521 agattacgct tacccatacg atgttccaga ttacgcttac ccatacgatg ttccagatta
     2581 cgcttgaggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
     2641 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2701 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2761 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2821 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2881 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2941 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3001 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3061 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3121 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3181 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3241 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3301 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3361 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3421 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3661 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3721 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3781 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4621 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4681 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4741 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4801 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4861 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4921 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4981 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5041 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5101 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5161 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5221 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5281 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5341 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5401 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5461 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5521 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5581 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5641 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5701 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5761 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5821 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5881 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5941 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6001 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6061 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6121 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6181 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6241 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6301 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6361 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6421 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6481 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6541 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6601 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6661 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6721 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6781 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6841 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6901 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6961 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7021 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7081 ccgaaaagtg ccacctgacg tc
//
