LOCUS       Exported                8503 bp ds-DNA     circular SYN 24-MAY-2025
DEFINITION  Mammalian Expression of tgRFPt-Micro.
ACCESSION   .
VERSION     .
KEYWORDS    pLL7.0: CAGEc-RFP-NLS (Nrdj1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8503)
  AUTHORS   Guntas G, Hallett RA, Zimmerman SP, Williams T, Yumerefendi H, Bear 
            JE, Kuhlman B
  TITLE     Engineering an improved light-induced dimer (iLID) for controlling 
            the localization and activity of signaling proteins.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Jan 6;112(1):112-7. doi: 
            10.1073/pnas.1417910112. Epub 2014 Dec 22.
  PUBMED    25535392
REFERENCE   2  (bases 1 to 8503)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 17, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8503
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2618..2931
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2850..2870
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2948..2964
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3006..3039
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     enhancer        3155..3458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3459..3662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3702..3711
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    5301..5334
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5390..5980
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5443..5463)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5861..5872)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5983..5999)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6509..6689
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6751..7339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6831..6850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7510..8370)
                     /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     8133..8152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8371..8475)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccgcg ctctagagat ccgacgccgc catctctagg
     2641 cccgcgccgg ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt
     2701 taatttgcat ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat
     2761 aatctgttct ttttaatact agctacattt tacatgatag gcttggattt ctataagaga
     2821 tacaaatact aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa
     2881 agtaattgtg tgttttgaga ctataaatat cccttggaga aaagccttgt taacgcgcgg
     2941 tgaccctcga ggtcgacggt atcgataagc tcgcttcacg agattccagc aggtcgaggg
     3001 acctaataac ttcgtatagc atacattata cgaagttata ttaagggttc caagcttaag
     3061 cggccgcgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     3121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     3181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     3241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     3301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     3361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     3421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     3481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     3541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     3601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     3661 ctggtttagt gaaccgtcag atccgctaga attgaccggt cgccaccatg ggaagagtag
     3721 cgatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt ttgtactttg
     3781 gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct atgatggaac
     3841 ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga gatttaatgg
     3901 aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac ctcctccaag
     3961 cctgggacct ctattatcat gtgttccgac gaatctcagg aaatggaaat ggaaacgctg
     4021 agttggttga actcgaaatt aagaccctac atggtgtgaa aacgatcagg tgtacccccg
     4081 accacccagt ttacactaaa aacaggggct acgtgcgagc ggatgagtta accgatgatg
     4141 atgagcttgt tgtagccatc gagaacctgt atggcgatga agtagatggc tattttcagg
     4201 gcatggaagc gaagacgtat ataggaaagc ttaaatcccg aaaaattgta tcaaacgagg
     4261 acacgtatga tattcagacg tcaactcata acttttttgc gaatgatata cttgttcata
     4321 atagtgagat cgtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
     4381 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     4441 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     4501 ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc
     4561 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
     4621 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     4681 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     4741 agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc
     4801 tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     4861 tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     4921 atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     4981 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatg
     5041 gcatggacga gctgtacaag tacccgtacg acgttccgga ctacgcttct ggaggtagtg
     5101 cggccgctcc aattgccgga tccaaggcta gcccgaaaaa gaaacgcaaa gttgggcgcg
     5161 gttctggttc agctgctgcc aagaagaaga agttggactc agggtcaggt tcttcaggtc
     5221 ctgctgccaa gagggtcaag ttggacgcct aaggatccac cggatcaaga taacctgcag
     5281 gcaattcgtc gagggaccta ataacttcgt atagcataca ttatacgaag ttatacatgt
     5341 ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct
     5401 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5461 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5521 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5581 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5641 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5701 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5761 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5821 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5881 ccttccttcc cgcgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5941 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     6001 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     6061 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     6121 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6181 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6241 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     6301 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     6361 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     6421 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     6481 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     6541 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6601 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6661 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
     6721 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6781 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6841 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6901 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6961 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7021 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7081 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7141 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7201 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7261 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7321 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7381 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7441 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7501 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7561 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7621 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7681 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7741 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7801 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7861 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7921 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7981 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8041 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8101 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8161 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8221 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8281 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8341 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8401 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8461 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
