LOCUS       sequence-241912-        4490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Nuclear Localized tagRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4490)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 4490)
  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"
COMMENT     Sequence Label: pAS2-his-24::tagRFP
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..74
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     66..83
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     67..83
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     93..112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             815..1525
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(1769..1789)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1769..1787)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1808..1824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1808..1824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1821..1843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1832..1848
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1856..1862,1863..1880,1881..1886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1901..1922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2039..2056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2210..2798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2290..2309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2969..3760,3761..3829))
                     /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     3592..3611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3830..3934)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3960..4415)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4097..4118)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4309..4328
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
       61 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
      121 agctccaccg cggactagtg gatcccccgg gctgcagcct aggatgtctg attccgctgt
      181 tgttgccgcc gctgtcgagc caaaggtccc aaaggctaag gccgccaagg ccgccaagcc
      241 agccgccaag gccgccaagc caaccaaagt tgccaaggcc aaggctccag tcgctcaccc
      301 accatacatc aacatgatca aggaggccat caagcagctc aaggaccgca aaggagcttc
      361 caagcaggca atcctcaagt tcatctccca aaactacaag ctcggagaca atgtcatcca
      421 gatcaatgct catctccgtc aggctctcaa gcgtggtgtc accagcaagg cccttgttca
      481 agccgccgga tccggagcca acggacgttt ccgtgtgcca gagaaggccg ccgccgccaa
      541 gaagccagca gcagccaaga agccagcagc agccaagaaa ccagcagccg ccaagaaagc
      601 cactggagag aagaaggcaa agaagccagc cgctgccaag ccaaagaagg ccgccaccgg
      661 agataagaag gtcaagaagg ccaagagccc aaagaaggtt gccaagccag ctgccaagaa
      721 ggtcgccaag tctccagcca agaaggccgc tccaaagaag atcgccaagc cagctgccaa
      781 gaaggccgcc aagccagccg ccaaggccgc atgcatggtg tctaagggcg aagagctgat
      841 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
      901 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
      961 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     1021 cagcagaacc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     1081 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     1141 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     1201 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga
     1261 gatgctgtac cccgctgacg gcggcctgga aggcagaacc gacatggccc tgaagctcgt
     1321 gggcgggggc cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa
     1381 gaacctcaag atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc
     1441 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     1501 tagcaaactg gggcacaaac ttaattccgg actcagatct cgagctcaag cttcgaattc
     1561 caactgaaag ctttggaatt cgctagccgg ccatacaagt aatccggatg atcgacgcca
     1621 acgtcgttga attttcaaat tttaaatact gaatatttgt tttttttcct attatttatt
     1681 tattctcttt gtgttttttt tcttgctttc taaaaaatta attcaatcca aatctaaaca
     1741 tgaggggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     1801 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1861 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcacccaa
     2101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3001 cagcgatctg cctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     3961 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4021 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4081 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4141 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4201 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4261 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4321 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4381 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     4441 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
//
