LOCUS       sequence-426914-        4179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for in vitro transcription of Vpx (SIV)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4179)
  AUTHORS   Levesque S, Cosentino A, Verma A, Genovese P, Bauer DE
  TITLE     Enhancing prime editing in hematopoietic stem and progenitor cells 
            by modulating nucleotide metabolism.
  JOURNAL   Nat Biotechnol. 2024 May 28. doi: 10.1038/s41587-024-02266-4.
  PUBMED    38806736
REFERENCE   2  (bases 1 to 4179)
  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 
            Biotechnol. 2024 May 28. doi: 10.1038/s41587-024-02266-4."
COMMENT     Sequence Label: T7_CC_Vpx-SIV_WPRE_IVT_Template
FEATURES             Location/Qualifiers
     source          1..4179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    686..1274
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(739..759)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1157..1168)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1277..1293)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1404..1420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1404..1420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1417..1439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1428..1444
                     /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(1452..1458,1459..1476,1477..1482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1497..1518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1635..1652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1806..2394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1886..1905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2565..3356,3357..3425))
                     /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     3188..3207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3426..3530)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3609..3628)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3800..4179
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatctcgag ctcggtacct
      241 aatacgacac actataaggg actcactatt tgttttcgcg cccagttgca aaaagtgtcg
      301 cttaagtcta gagccaccat gtcagatccc agggagagaa tcccacctgg aaacagtgga
      361 gaagagacaa taggagaggc cttcgaatgg ctaaacagaa cagtagagga gataaacaga
      421 gaggcagtaa accacctacc aagggagctg attttccagg tttggcaaag gtcttgggaa
      481 tactggcatg atgaacaagg gatgtcacaa agctatgtaa aatacagata cttgtgttta
      541 atgcaaaagg ctttatttat gcattgcaag aaaggctgta gatgtctagg ggaaggacac
      601 ggggcaggag gatggagacc aggacctcct cctcctcccc ctccaggact agcatgataa
      661 cccgggaccg gtcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
      721 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
      781 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
      841 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
      901 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
      961 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     1021 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     1081 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     1141 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     1201 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     1261 ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta gaaaacatgc tggggatgcg
     1321 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct
     1381 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1441 acaattccac acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga
     1501 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1561 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2401 cctttgatct tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt
     2461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2701 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2761 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2821 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2881 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2941 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3001 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3061 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3121 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3181 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3241 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3301 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3361 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3421 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3541 cgaaaagtgc cacctgacgt cgacggatcg ggagatcgat ctcccgatcc cctagggtct
     3601 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     3661 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     3721 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     3781 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     3841 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     3901 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     3961 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     4021 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     4081 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     4141 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
