LOCUS       sequence-258154-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIRTS-8 (bdefensin 3-TBP6.7-hADAR(299-701)E488Q) in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5576)
  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 5576)
  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-8: bdefensin 3-TBP6.7-hADAR(299-701)E488Q
FEATURES             Location/Qualifiers
     source          1..5576
                     /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             2645..2692
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2699..2722
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2840..2858)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2895..3016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2906..2925)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2960..2979
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3415..4209
                     /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(3469..3488)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4079..4098
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4381..4969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4461..4480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5085..5368
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        join(5559..5576,1..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 gggcggtagt ggaggaagcg gggggagcgg agggagtggt ggatccggcg gctccttgca
     1441 cttggatcaa acgccatctc gccagcctat tcccagtgag ggtcttcagc tgcatttacc
     1501 gcaggtttta gctgacgctg tctcacgcct ggtcctgggt aagtttggtg acctgaccga
     1561 caacttctcc tcccctcacg ctcgcagaaa agtgctggct ggagtcgtca tgacaacagg
     1621 cacagatgtt aaagatgcca aggtgataag tgtttctaca ggaacaaaat gtattaatgg
     1681 tgaatacatg agtgatcgtg gccttgcatt aaatgactgc catgcagaaa taatatctcg
     1741 gagatccttg ctcagatttc tttatacaca acttgagctt tacttaaata acaaagatga
     1801 tcaaaaaaga tccatctttc agaaatcaga gcgagggggg tttaggctga aggagaatgt
     1861 ccagtttcat ctgtacatca gcacctctcc ctgtggagat gccagaatct tctcaccaca
     1921 tgagccaatc ctggaagaac cagcagatag acacccaaat cgtaaagcaa gaggacagct
     1981 acggaccaaa atagagtctg gtcaggggac gattccagtg cgctccaatg cgagcatcca
     2041 aacgtgggac ggggtgctgc aaggggagcg gctgctcacc atgtcctgca gtgacaagat
     2101 tgcacgctgg aacgtggtgg gcatccaggg atccctgctc agcattttcg tggagcccat
     2161 ttacttctcg agcatcatcc tgggcagcct ttaccacggg gaccaccttt ccagggccat
     2221 gtaccagcgg atctccaaca tagaggacct gccacctctc tacaccctca acaagccttt
     2281 gctcagtggc atcagcaatg cagaagcacg gcagccaggg aaggccccca acttcagtgt
     2341 caactggacg gtaggcgact ccgctattga ggtcatcaac gccacgactg ggaaggatga
     2401 gctgggccgc gcgtcccgcc tgtgtaagca cgcgttgtac tgtcgctgga tgcgtgtgca
     2461 cggcaaggtt ccctcccact tactacgctc caagattacc aaacccaacg tgtaccatga
     2521 gtccaagctg gcggcaaagg agtaccaggc cgccaaggcg cgtctgttca cagccttcat
     2581 caaggcgggg ctgggggcct gggtggagaa gcccaccgag caggaccagt tctcactcac
     2641 gcccaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agggatccga
     2701 ttacaaggac gacgatgaca agtagtgttt aaacgaattc gggctcggta cacggggatc
     2761 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     2821 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     2881 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2941 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3001 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3061 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaattcta
     3121 atggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     3181 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
     3241 cttgcgccct gagtgcttgc ggcagcgtga gcttcaaaag aattgccagc tggggcgccc
     3301 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     3361 tgatggcgca ggggatcaag atctgatcaa gagacaggat gacggtcgtt tcgcatgctt
     3421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3841 cgagcacgca ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgtatgcc ggatggtgag
     3961 gatctcgtcg tgactcatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4201 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aaccggtatc
     4261 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4321 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4381 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4441 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4501 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4561 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4621 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4681 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4741 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4801 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4861 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4921 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     4981 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5041 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     5101 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     5161 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     5221 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     5281 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     5341 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     5401 cggcgtagag gatcgagatc ttcaatattg gccattagcc atattattca ttggttatat
     5461 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     5521 catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat tgacta
//
