LOCUS       pT3TS-RfxCas13d_        6154 bp    DNA     circular SYN 06-MAY-2025
DEFINITION  2C-NLS.
ACCESSION   2C-NLS
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.2
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
                     /note="color: #ffff00"
     CDS             772..864
                     /codon_start=1
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.6"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="APNTQTASPRALADSLMQLARQVSRLESGQ"
     protein_bind    877..898
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
                     /note="color: #31849b"
     promoter        913..943
                     /label=lac promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
                     /note="color: #ffffff"
     protein_bind    951..967
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
                     /note="color: #31849b"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
                     /note="color: #ffffff"
     5'UTR           1055..1097
                     /label=Xenopus globin 5'-UTR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=5'UTR"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1113..1122
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             1119..4022
                     /codon_start=1
                     /label=RfxCas13d
                     /note="color: #993366"
                     /translation="MAIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGD
                     SIRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETL
                     EKRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFST
                     VYTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYI
                     INYGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRIT
                     NELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDR
                     KDMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIF
                     VINLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILP
                     AGRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYA
                     FFKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLD
                     ENGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADI
                     QKKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGR
                     ENAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKL
                     EEKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEK
                     KAEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYV
                     HAYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLC
                     VPFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4032..4052
                     /codon_start=1
                     /label=SV40-NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             4065..4112
                     /codon_start=1
                     /label=nucleoplasmin NLS
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=97.9"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="KRSAATKKAGQAKKKK"
     3'UTR           4137..4262
                     /label=Xenopus globin 3'-UTR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=3'UTR"
                     /note="color: #a6acb3"
     promoter        complement(4354..4372)
                     /label=T7 promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
                     /note="color: #ffffff; direction: LEFT"
     CDS             4375..4532
                     /codon_start=1
                     /label=lacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=90.8"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="IHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPN
                     SCAA*MA"
     rep_origin      complement(4564..4992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
                     /note="color: #ffff00"
     promoter        5019..5123
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
                     /note="color: #ffffff"
     CDS             5124..5984
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /note="color: #ccffcc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tggagctaag cttgcttgtt ctttttgcag aagctcagaa
     1081 taaacgctca actttggcag atctaactcg acgccaccat ggcgatcgaa aaaaaaaagt
     1141 ccttcgccaa gggcatgggc gtgaagtcca cactcgtgtc cggctccaaa gtgtacatga
     1201 caaccttcgc cgaaggcagc gacgccaggc tggaaaagat cgtggagggc gacagcatca
     1261 ggagcgtgaa tgagggcgag gccttcagcg ctgaaatggc cgataaaaac gccggctata
     1321 agatcggcaa cgccaaattc agccatccta agggctacgc cgtggtggct aacaaccctc
     1381 tgtatacagg acccgtccag caggatatgc tcggcctgaa ggaaactctg gaaaagaggt
     1441 acttcggcga gagcgctgat ggcaatgaca atatttgtat ccaggtgatc cataacatcc
     1501 tggacattga aaaaatcctc gccgaataca ttaccaacgc cgcctacgcc gtcaacaata
     1561 tctccggcct ggataaggac attattggat tcggcaagtt ctccacagtg tatacctacg
     1621 acgaattcaa agaccccgag caccataggg ccgctttcaa caataacgat aagctcatca
     1681 acgccatcaa ggcccagtat gacgagttcg acaacttcct cgataacccc agactcggct
     1741 atttcggcca ggcctttttc agcaaggagg gcagaaatta catcatcaat tacggcaacg
     1801 aatgctatga cattctggcc ctcctgagcg gactgaggca ctgggtggtc cataacaacg
     1861 aagaagagtc caggatctcc aggacctggc tctacaacct cgataagaac ctcgacaacg
     1921 aatacatctc caccctcaac tacctctacg acaggatcac caatgagctg accaactcct
     1981 tctccaagaa ctccgccgcc aacgtgaact atattgccga aactctggga atcaaccctg
     2041 ccgaattcgc cgaacaatat ttcagattca gcattatgaa agagcagaaa aacctcggat
     2101 tcaatatcac caagctcagg gaagtgatgc tggacaggaa ggatatgtcc gagatcagga
     2161 aaaatcataa ggtgttcgac tccatcagga ccaaggtcta caccatgatg gactttgtga
     2221 tttataggta ttacatcgaa gaggatgcca aggtggctgc cgccaataag tccctccccg
     2281 ataatgagaa gtccctgagc gagaaggata tctttgtgat taacctgagg ggctccttca
     2341 acgacgacca gaaggatgcc ctctactacg atgaagctaa tagaatttgg agaaagctcg
     2401 aaaatatcat gcacaacatc aaggaattta ggggaaacaa gacaagagag tataagaaga
     2461 aggacgcccc tagactgccc agaatcctgc ccgctggccg tgatgtttcc gccttcagca
     2521 aactcatgta tgccctgacc atgttcctgg atggcaagga gatcaacgac ctcctgacca
     2581 ccctgattaa taaattcgat aacatccaga gcttcctgaa ggtgatgcct ctcatcggag
     2641 tcaacgctaa gttcgtggag gaatacgcct ttttcaaaga ctccgccaag atcgccgatg
     2701 agctgaggct gatcaagtcc ttcgctagaa tgggagaacc tattgccgat gccaggaggg
     2761 ccatgtatat cgacgccatc cgtattttag gaaccaacct gtcctatgat gagctcaagg
     2821 ccctcgccga caccttttcc ctggacgaga acggaaacaa gctcaagaaa ggcaagcacg
     2881 gcatgagaaa tttcattatt aataacgtga tcagcaataa aaggttccac tacctgatca
     2941 gatacggtga tcctgcccac ctccatgaga tcgccaaaaa cgaggccgtg gtgaagttcg
     3001 tgctcggcag gatcgctgac atccagaaaa aacagggcca gaacggcaag aaccagatcg
     3061 acaggtacta cgaaacttgt atcggaaagg ataagggcaa gagcgtgagc gaaaaggtgg
     3121 acgctctcac aaagatcatc accggaatga actacgacca attcgacaag aaaaggagcg
     3181 tcattgagga caccggcagg gaaaacgccg agagggagaa gtttaaaaag atcatcagcc
     3241 tgtacctcac cgtgatctac cacatcctca agaatattgt caatatcaac gccaggtacg
     3301 tcatcggatt ccattgcgtc gagcgtgatg ctcaactgta caaggagaaa ggctacgaca
     3361 tcaatctcaa gaaactggaa gagaagggat tcagctccgt caccaagctc tgcgctggca
     3421 ttgatgaaac tgcccccgat aagagaaagg acgtggaaaa ggagatggct gaaagagcca
     3481 aggagagcat tgacagcctc gagagcgcca accccaagct gtatgccaat tacatcaaat
     3541 acagcgacga gaagaaagcc gaggagttca ccaggcagat taacagggag aaggccaaaa
     3601 ccgccctgaa cgcctacctg aggaacacca agtggaatgt gatcatcagg gaggacctcc
     3661 tgagaattga caacaagaca tgtaccctgt tcagaaacaa ggccgtccac ctggaagtgg
     3721 ccaggtatgt ccacgcctat atcaacgaca ttgccgaggt caattcctac ttccaactgt
     3781 accattacat catgcagaga attatcatga atgagaggta cgagaaaagc agcggaaagg
     3841 tgtccgagta cttcgacgct gtgaatgacg agaagaagta caacgatagg ctcctgaaac
     3901 tgctgtgtgt gcctttcggc tactgtatcc ccaggtttaa gaacctgagc atcgaggccc
     3961 tgttcgatag gaacgaggcc gccaagttcg acaaggagaa aaagaaggtg tccggcaatt
     4021 ccggatccgg acctaagaaa aagaggaagg tgggcagcgg aagcaaaagg tcggcggcca
     4081 cgaaaaaggc cggccaggca aaaaagaaaa agtgaccgcg gatctggtta ccactaaacc
     4141 agcctcaaga acacccgaat ggagtctcta agctacataa taccaactta cactttacaa
     4201 aatgttgtcc cccaaaatgt agccattcgt atctgctcct aataaaaaga aagtttcttc
     4261 acattctaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaacc cccccccccc ccccctgcag
     4321 ggccaagtcg gcctctagag gtacccaatt cgccctatag tgagtcgtat tacaattcac
     4381 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     4441 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     4501 cttcccaaca gttgcgcagc ctgaatggcg aatggcaaat tgtaagcgtt aatattttgt
     4561 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4621 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4681 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4741 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4801 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4861 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4921 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4981 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5041 ttttctaaat acattcaaat atgtatctgc tcatgagaca ataaccctga taaatgcttc
     5101 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5161 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5221 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5281 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5341 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5401 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5461 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5521 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5581 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5641 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5701 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5761 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5821 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5881 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5941 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6001 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6061 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6121 cgtcagaccc cgtagaaaag atcaaaggat cttc
//
