LOCUS       sequence-403018-        3727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For production of template RNA for use in PRINT, mCherry under CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3727)
  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 3727)
  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: TCARZ-CMV-Cherry-PAmin-GF3-R4A22
FEATURES             Location/Qualifiers
     source          1..3727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1199..1218
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1199..1217
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        1332..1635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1636..1839
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1790..1810
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1960..1969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      complement(3139..3727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3219..3238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtccca atgatgtccc aatgatctta aggaattcta
     1201 atacgactca ctatagggta aacggcggga gtaactatga ctctcttaag gagtcataga
     1261 gccagaacct cctcgtggtc ccgctgggca cagggattaa tttttctgtg gcaaatttga
     1321 ctggctctag acgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1381 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1441 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     1501 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     1561 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     1621 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg ataggggttt
     1681 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     1741 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     1801 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgcttgtct
     1861 cgctccgggg aacgctcgga aactcccggc cgccgccacc cgcgtctgtt ctgttacaca
     1921 agggaagaaa agccgctgcc gcactccgag tgtcgtacgg ccaccatggt gagcaaaggc
     1981 gaagaagata acatggcgat tattaaagaa tttatgcgct ttaaagtgca tatggaaggc
     2041 agcgtgaacg gccatgaatt tgaaattgaa ggcgaaggcg aaggccgccc gtatgaaggc
     2101 acccagaccg cgaaactgaa agtgaccaaa ggcggcccgc tgccgtttgc gtgggatatt
     2161 ctgagcccgc agtttatgta tggcagcaaa gcgtatgtga aacatccggc ggatattccg
     2221 gattatctga aactgagctt tccggaaggc tttaaatggg aacgcgtgat gaactttgaa
     2281 gatggcggcg tggtgaccgt gacccaggat agcagcctgc aggatggcga atttatttat
     2341 aaagtgaaac tgcgcggcac caactttccg agcgatggcc cggtgatgca gaaaaaaacc
     2401 atgggctggg aagcgagcag cgaacgcatg tatccggaag atggcgcgct gaaaggcgaa
     2461 attaaacagc gcctgaaact gaaagatggc ggccattatg atgcggaagt gaaaaccacc
     2521 tataaagcga aaaaaccggt gcagctgccg ggcgcgtata acgtgaacat taaactggat
     2581 attaccagcc ataacgaaga ttataccatt gtggaacagt atgaacgcgc ggaaggccgc
     2641 catagcaccg gcggcatgga tgaactgtat aaatagtgaa agcttaataa aagatcttta
     2701 tgttcattag atctgtgtgt tggtttttgt gtggcggccg ctagggtaga taatctttgt
     2761 atagtggggg gggatctcat gtaccgggtt tcttttattt gattttcaat aaaacagacg
     2821 gtagctaggt tcgcaaggca gccacaagcc aaagataggt agggtgctca tagtgagtag
     2881 ggacagtgcc ttttgattca caacgcgtca ataccatctg acacggatac ccttaccgga
     2941 cttgtcatga tctcccagac ttgtccaagg tggacgggcc acctttactt aacccggaaa
     3001 aggaacatat attaattata tgtgttcgga aaatagcaaa aaaaaaaaaa aaaaaaaaaa
     3061 agtcttcatc ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3541 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3721 atctcaa
//
