LOCUS       sequence-258153-        4931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIRTS-6 (bdefensin 3-TBP6.7-YTHDF2(1-200)) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4931)
  AUTHORS   Rauch S, He E, Srienc M, Zhou H, Zhang Z, Dickinson BC
  TITLE     Programmable RNA-Guided RNA Effector Proteins Built from Human 
            Parts.
  JOURNAL   Cell. 2019 Jun 27;178(1):122-134.e12. doi: 
            10.1016/j.cell.2019.05.049. Epub 2019 Jun 20.
  PUBMED    31230714
REFERENCE   2  (bases 1 to 4931)
  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.cell.2019.05.049"; journalName: "Cell"; date: 
            "2019-06-27- 27"; volume: "178"; issue: "1"; pages: "122-134.e12"
COMMENT     Sequence Label: CIRTS-6: bdefensin 3-TBP6.7-YTHDF2(1-200)
FEATURES             Location/Qualifiers
     source          1..4931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          702..834
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     817..836
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      878..887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1394..1420
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2054..2077
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2195..2213)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2250..2371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2261..2280)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2315..2334
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2770..3564
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2824..2843)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3434..3453
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3736..4324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3816..3835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4440..4723
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        join(4914..4931,1..362)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcact agaagcttta
      601 ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct
      661 cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
      721 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
      781 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
      841 actcccagtt caattacagc tcttaaggct agagtatgcc accatgggaa ttatcaacac
      901 ccttcaaaaa tactactgtc gcgtccgtgg gggtcgttgc gcagtcctgt cgtgcctgcc
      961 taaagaggag cagatcggta aatgctcaac ccgtggacgt aagtgctgtc gtcgcaagaa
     1021 aggcggttct ggaggctcag gggggagcgg agggtccggt ggatcaggag gctccatggc
     1081 agttcccgag acccgcccta accacactat ttatatcaac aacctcaata gcaagatcaa
     1141 gaaggatgag ctaaaaaagt ccctgtacgc catcttctcc cagtttggcc agatcctgga
     1201 tatcctggta ccgcggcagc gtaccccgag gggccaggcc tttgtcatct tcaaggaggt
     1261 cagcagcgcc accaacgccc tgcgctccat gcagggtttc cctttctatg acaaacctat
     1321 gcgtatccag tatgccaaga ccgacaaacg tatcccggcc aagatgaaag gcaccttcgt
     1381 gggatccctt caactgcctc cacttgaaag actgacactg ggcggtagtg gaggaagcgg
     1441 ggggagcgga gggagtggtg gatccggcgg ctccatgtcg gccagcagcc tcttggagca
     1501 gagaccaaaa ggtcaaggaa acaaagtaca aaatggatct gtacatcaaa aggatggatt
     1561 aaacgatgat gattttgaac cttacttgag tccacaggca aggcccaata atgcatatac
     1621 tgccatgtca gattcctact tacccagtta ctacagtccc tccattggct tctcctattc
     1681 tttgggtgaa gctgcttggt ctacgggggg tgacacagcc atgccctact taacttctta
     1741 tggacagctg agcaacggag agccccactt cctaccagat gcaatgtttg ggcaaccagg
     1801 agccctaggt agcactccat ttcttggtca gcatggtttt aatttctttc ccagtgggat
     1861 tgacttctca gcatggggaa ataacagttc tcagggacag tctactcaga gctctggata
     1921 tagtagcaat tatgcttatg cacctagctc cttaggtgga gccatgattg atggacagtc
     1981 agcttttgcc aatgagaccc tcaataaggc tcctggcatg aatactatag accaagggat
     2041 ggcagcagga tccgattaca aggacgacga tgacaagtag tgtttaaacg aattcgggct
     2101 cggtacacgg ggatcctcta gagtcgacct gcaggcatgc aagctgatcc ggctgctaac
     2161 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2221 cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2281 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2341 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2401 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     2461 taaaatcgaa ttctaatgga tcctctttgc gcttgcgttt tcccttgtcc agatagccca
     2521 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg
     2581 cttcctttag cagcccttgc gccctgagtg cttgcggcag cgtgagcttc aaaagaattg
     2641 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     2701 ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgacgg
     2761 tcgtttcgca tgcttgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2821 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2881 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2941 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3001 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3061 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3121 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3181 aaacatcgca tcgagcgagc acgcactcgg atggaagccg gtcttgtcga tcaggatgat
     3241 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgt
     3301 atgccggatg gtgaggatct cgtcgtgact catggcgatg cctgcttgcc gaatatcatg
     3361 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3421 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3481 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3541 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3601 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4141 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     4441 cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat
     4501 ggcagcttca gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat
     4561 tcatacggtt aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat
     4621 ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg
     4681 gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc
     4741 cggccacgat gcgtccggcg tagaggatcg agatcttcaa tattggccat tagccatatt
     4801 attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     4861 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttggcattg
     4921 attattgact a
//
