LOCUS       sequence-472668-        5876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For production of C-term flag-tagged TaGu RT-dead ORF mRNA for mRNA 
            transfection in PRINT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5876)
  AUTHORS   Zhang X, Van Treeck B, Horton CA, McIntyre JJR, Palm SM, Shumate JL,
            Collins K
  TITLE     Harnessing eukaryotic retroelement proteins for transgene insertion 
            into human safe-harbor loci.
  JOURNAL   Nat Biotechnol. 2024 Feb 20. doi: 10.1038/s41587-024-02137-y.
  PUBMED    38379101
REFERENCE   2  (bases 1 to 5876)
  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 Feb 20. doi: 10.1038/s41587-024-02137-y."
COMMENT     Sequence Label: pT7-TaGu-RTD-3xF-ORF
FEATURES             Location/Qualifiers
     source          1..5876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      117..126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3684..3749
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     rep_origin      complement(4164..4752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4244..4263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4923..5714,5715..5783))
                     /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     5546..5565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5784..5876,1..12))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcccaatga ttaatacgac
       61 tcactataag gaataaacta gtattcttct ggtccccaca gactcagaga gaacccgcca
      121 ccatggagaa ggtgatggtc accgttccgg acaagaaccc tccatgtcca tgctgcggca
      181 cccgggtgaa cagcgtgctg aaccttatcg agcacctgaa ggtcagccac ggaaagcggg
      241 gcgtgtgctt ccggtgcgcg aagtgtggga aggaaaatag caactaccac agcgtggtgt
      301 gccatttccc aaaatgtaga ggacctgaga cagagaaagc tccagccggc gagtggattt
      361 gtgaagtgtg caacagagac ttcaccacca aaatcggcct gggccagcat aagagattag
      421 ctcaccccgc tgttcggaat caggagagaa tcgtcgccag ccagcctaag gaaactagca
      481 acagaggcgc tcacaaaaga tgctggacca aagaggagga ggaactgctg atcagactgg
      541 aagcccagtt tgagggcaac aagaacatca acaagctgat cgcagaacac atcaccacaa
      601 agaccgccaa gcagatctcc gataagagga gactgctgag ccgcaagcct gccgaggagc
      661 ctagagagga acctggcaca tgccaccaca ccagacgggc agctgccagc ctgagaacag
      721 agcccgagat gtcccaccac gctcaggccg aagatcggga taatggccct ggccggagac
      781 ccctgcctgg tagagccgcc gccggcggca gaacaatgga cgagatcagg agacaccctg
      841 acaagggcaa tggacagcag cggcctacca aacagaagtc tgaggagcag ctgcaagcct
      901 actacaaaaa gacactggag gagcggctga gcgccggagc cctgaatacc ttcccccgcg
      961 ccttcaagca agtgatggaa ggcagagata tcaagctggt gatcaaccag acagcccagg
     1021 actgcttcgg ctgtctggag agcatctctc agatcagaac agcgaccaga gataagaaag
     1081 ataccgtgac cagagagaag cacccgaaga agccattcca aaagtggatg aaggaccggg
     1141 ccataaagaa gggcaactac ctgaggttcc agagattgtt ctacctggac cgaggcaaac
     1201 tggccaagat catcctggac gatatcgaat gcctgagctg cgacatcccc ctgtctgaga
     1261 tctacagcgt gtttaagaca agatgggaaa ccacaggaag cttcaaatct ctgggcgatt
     1321 ttaaaaccta cggcaaggcc gacaacactg ccttcagaga actgattaca gctaaggaaa
     1381 tcgagaaaaa cgtgcaggag atgagcaagg gcagcgcccc tggccctgac ggtatcaccc
     1441 tgggagatgt ggtgaaaatg gaccccgagt tcagccgcac aatggagatc ttcaacctgt
     1501 ggctgaccac tggcaaaatc cctgacatgg tgcggggatg ccgcaccgtg ctgataccta
     1561 agagcagcaa gccagataga ctgaaggata tcaacaactg gcggcctatc acaatcggca
     1621 gcatcctgct ccggctgttt agcagaatcg taaccgcccg tctgagtaag gcctgtcctc
     1681 tgaaccccag acagcggggc ttcatccggg ccgccggctg cagcgagaac ctcaaactgc
     1741 tgcagaccat catctggtcg gccaaacgag agcacagacc tctcggcgtg gtgttcgtgg
     1801 acatcgccaa agcttttgat accgtctccc accagcacat catccacgcc ctgcagcagc
     1861 gggaagtgga ccctcacatc gtgggactgg tgtccaacat gtacgagaac atcagcactt
     1921 atatcaccac caagagaaac acccacacag acaagatcca gatcagagtg ggcgttaagc
     1981 agggcgatcc tatgtctcct ctgctgttca acctggctat ggaccctctg ctctgtaaac
     2041 tggaggagtc tggcaaggga taccacagag gccaaagctc catcaccgct atggcctttg
     2101 ccgccgcact ggtgctgctg tctgattctt gggagaacat gaataccaac ataagcatcc
     2161 tggaaacatt ctgcaacctt acaggcctga aaacccaggg acaaaagtgc cacggctttt
     2221 acatcaagcc taccaaagac agctacacca tcaacgactg cgccgcctgg accatcaacg
     2281 gcaccccttt aaacatgatc gaccctggag aaagcgagaa gtatctgggc ctgcagttcg
     2341 acccctggat cggcatcgct agaagcggcc tgagcacaaa gctggacttt tggctgcaga
     2401 gaattgacca ggcccctctg aagcctctgc agaaaaccga catcctgaaa acctacacaa
     2461 tccctcggct gatctacatt gccgatcaca gcgaggtaaa gaccgccctg ctggaaaccc
     2521 tggaccaaaa gatcagaacc gccgtgaagg aatggctcca cctgcccccc tgcacctgcg
     2581 acgctatcct gtacagcagc accagggacg gcggcctggg catcaccaag ctggcgggcc
     2641 tgatcccctc cgtccaggct agacggctgc atagaatcgc ccagagcagc gatgacacaa
     2701 tgaagtgctt tatggaaaag gaaaagatgg aacagctgca caagaagctg tggattcagg
     2761 ccggtggcga cagagagaac attcctagca tctgggaggc gccgcctagt agcgagcctc
     2821 ccaacaacgt gtctaccaat tctgagtggg aagcccctac acagaaggac aagttcccta
     2881 agccttgtaa ttggagaaag aacgagttca agaagtggac aaagctggcc tctcagggcc
     2941 ggggaattgt gaatttcgag cgggacaaga tcagcaatca ctggattcag tactacagaa
     3001 gaatcccaca cagaaagctg ctgacggccc ttcagctgag agccaacgtg taccccacgc
     3061 gggagttcct ggccagaggt agacaggacc agtacatcaa ggcctgcaga cattgtgatg
     3121 ctgatatcga gtcttgcgcc cacatcatcg gcaactgccc tgtgacacag gacgcgagaa
     3181 tcaaaagaca caactacatc tgcgagctgc tgctggaaga ggccaagaag aaggactggg
     3241 tggtgttcaa ggaaccccac atcagagaca gcaataagga actctataaa cctgacctga
     3301 tcttcgtgaa ggacgcccgg gccctggtcg tggacgtgac cgtgagatac gaggccgcca
     3361 agtctagcct ggaggaggct gccgccgaga aagtgcggaa gtacaagcac cttgaaacag
     3421 aagtgcggca cctgaccaac gccaaggacg tcacattcgt gggcttcccc ctgggcgcca
     3481 gaggcaaatg gcaccaggat aacttcaagc tgctgacaga gctgggcctg agcaagtccc
     3541 gccaagtgaa gatggccgag accttcagca ccgtggccct gttctcctct gtggacatcg
     3601 tgcacatgtt cgctagcaga gccagaaaga gcatggtgat gcatatgggt ggaggtagcg
     3661 ggggcagtgg agggatgggg agcgactaca aagaccatga cggtgattat aaagatcatg
     3721 acatcgatta caaggatgac gatgacaaga agtgatgacc tcgagctggt actgcatgca
     3781 cgcaatgcta gctgcccctt tcccgtcctg ggtaccccga gtctcccccg acctcgggtc
     3841 ccaggtatgc tcccacctcc acctgcccca ctcaccacct ctgctagttc cagacacctc
     3901 ccaagcacgc agcaatgcag ctcaaaacgc ttagcctagc cacaccccca cgggaaacag
     3961 cagtgattaa cctttagcaa taaacgaaag tttaactaag ctatactaac cccagggttg
     4021 gtcaatttcg tgccagccac accctggagc tagcaaaaaa aaaaaaaaaa aaaaaaaaaa
     4081 aaaaaagtct tcatcggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaat
//
