LOCUS       sequence-370603-        5310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HERV-Kcon RT (human codon optimization).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5310)
  AUTHORS   Grunewald J, Miller BR, Szalay RN, Cabeceiras PK, Woodilla CJ, Holtz
            EJB, Petri K, Joung JK
  TITLE     Engineered CRISPR prime editors with compact, untethered reverse 
            transcriptases.
  JOURNAL   Nat Biotechnol. 2023 Mar;41(3):337-343. doi: 
            10.1038/s41587-022-01473-1. Epub 2022 Sep 26.
  PUBMED    36163548
REFERENCE   2  (bases 1 to 5310)
  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: 
            "10.1038/s41587-022-01473-1"; journalName: "Nat Biotechnol"; date: 
            "2023-03"; volume: "41"; issue: "3"; pages: "337-343"
COMMENT     Sequence Label: HERV-Kcon-RT for Split-PE (pJUL3120)
FEATURES             Location/Qualifiers
     source          1..5310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             701..721
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2552..2572
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2581..2598
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2621..2638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2627..2851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2922..2938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2922..2938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2935..2957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2946..2962
                     /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(2970..2976,2977..2994,2995..3000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3015..3036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3153..3170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3324..3912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3404..3423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4083..4874,4875..4943))
                     /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     4706..4725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4944..5048)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5127..5146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      601 aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag ggagagccgc
      661 caccatgaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga agcggaaagt
      721 caaaagcaga aaacggagaa atagagtgtc cttcctgggc gctgccacag tggaaccacc
      781 taagcccatc cctctgacat ggaaaacaga gaagcctgtg tgggtcaacc agtggcctct
      841 gcctaagcag aagctggagg ctctccacct gctggccaac gagcagcttg agaagggcca
      901 catcgagccc agctttagcc cttggaacag ccctgtgttc gtgatccaga agaagagcgg
      961 caagtggcgg atgctgacag atctgagagc tgtgaacgcc gtgatccaac ccatgggccc
     1021 cctgcagcca ggcctgcctt cccctgctat gatccctaaa gattggcctc tgatcatcat
     1081 cgacctgaaa gactgcttct tcacaatccc actcgccgag caggattgcg agaagttcgc
     1141 cttcaccatc cccgccatca acaacaagga gcctgccacc agattccagt ggaaggtgct
     1201 gcctcagggc atgctgaatt ctccaacaat ctgccagacc ttcgtgggca gagctctgca
     1261 gcctgttaga gaaaaattca gcgactgcta catcattcac tacatcgatg acatcctgtg
     1321 cgccgctgaa accaaggata agttgatcga ctgttacacc ttcctgcaag ccgaggtggc
     1381 caatgccgga ctggctatcg cctctgataa gatccagacc agcacacctt tccactacct
     1441 gggcatgcag atcgagaacc ggaagatcaa gccacagaaa atcgagatca gaaaggacac
     1501 cctgaagacc ctgaacgact tccagaaact cctgggggat atcaactgga tcagacctac
     1561 cctgggaatc cctacgtacg ccatgagcaa cctgttcagc atcctgaggg gcgacagcga
     1621 cctgaacagc aagagaatgc tgacccctga ggccacaaaa gagatcaagc tggtggaaga
     1681 gaagatccag tctgctcaaa tcaacagaat cgatcccctg gcccctcttc agttgctgat
     1741 tttcgccact gcccatagcc ccaccggcat tatcatccag aacaccgacc tggtggaatg
     1801 gtcttttctg ccccacagca ccgtgaagac atttacactg tacctggacc agatcgccac
     1861 cctgatcggc caaacaagac tgcggatcat caagctgtgt ggcaacgacc ccgacaagat
     1921 cgtggtgcct ctgaccaagg aacaggtgcg gcaggctttt attaactccg gcgcctggca
     1981 gatcggactg gccaacttcg ttggcatcat cgacaatcac tatcctaaga ccaagatctt
     2041 ccaatttctg aagctgacca cctggattct gcctaagatt acaagacggg aacccctgga
     2101 gaacgccctg accgtgttca ccgacggatc ttccaacggc aaagccgcct acaccggccc
     2161 taaggaaaga gtgattaaga caccatacca gagcgcccag agagccgaac tggtcgccgt
     2221 gatcaccgtg ctgcaggact tcgaccagcc tatcaatatc atcagcgaca gtgcctatgt
     2281 ggtgcaggcc acccgggacg tggaaaccgc cctgatcaag tacagcatgg acgatcagct
     2341 caaccagctg tttaacctgc tgcagcagac cgtgcggaag agaaacttcc ccttctacat
     2401 cacccacatc cgcgcccaca ccaacctgcc cggccctctg acaaaggcca atgagcaggc
     2461 tgatctgctg gtgtctagcg ccctgattaa ggcccaggag ctgcacgcct ctggcggctc
     2521 aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag tctaaccggt
     2581 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     2641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2701 cccactgtcc tttcctaata aaatgagaaa attgcatcgc attgtctgag taggtgtcat
     2761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2821 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2881 tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3001 gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacactca gtggaacgaa
     3961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5041 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct
     5101 cccgatcccc tagggtctac tctcagtaca atctgctctg atgccgcata gttaagccag
     5161 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     5221 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     5281 gcgctgcttc gcgatgtacg ggccagatat
//
