LOCUS       sequence-237332-        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"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1348..2058
                     /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(2302..2322)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2302..2320)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2341..2357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2341..2357)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2354..2376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2365..2381
                     /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(2389..2395,2396..2413,2414..2419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2434..2455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2572..2589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2743..3331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2823..2842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3502..4293,4294..4362))
                     /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     4125..4144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4363..4467)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggacta gtggatcccc cgggctgcag cctaggatgt ctgattccgc tgttgttgcc
      721 gccgctgtcg agccaaaggt cccaaaggct aaggccgcca aggccgccaa gccagccgcc
      781 aaggccgcca agccaaccaa agttgccaag gccaaggctc cagtcgctca cccaccatac
      841 atcaacatga tcaaggaggc catcaagcag ctcaaggacc gcaaaggagc ttccaagcag
      901 gcaatcctca agttcatctc ccaaaactac aagctcggag acaatgtcat ccagatcaat
      961 gctcatctcc gtcaggctct caagcgtggt gtcaccagca aggcccttgt tcaagccgcc
     1021 ggatccggag ccaacggacg tttccgtgtg ccagagaagg ccgccgccgc caagaagcca
     1081 gcagcagcca agaagccagc agcagccaag aaaccagcag ccgccaagaa agccactgga
     1141 gagaagaagg caaagaagcc agccgctgcc aagccaaaga aggccgccac cggagataag
     1201 aaggtcaaga aggccaagag cccaaagaag gttgccaagc cagctgccaa gaaggtcgcc
     1261 aagtctccag ccaagaaggc cgctccaaag aagatcgcca agccagctgc caagaaggcc
     1321 gccaagccag ccgccaaggc cgcatgcatg gtgtctaagg gcgaagagct gattaaggag
     1381 aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca
     1441 tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag
     1501 ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaga
     1561 accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt ccctgagggc
     1621 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac
     1681 accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca
     1741 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccaacac cgagatgctg
     1801 taccccgctg acggcggcct ggaaggcaga accgacatgg ccctgaagct cgtgggcggg
     1861 ggccacctga tctgcaactt caagaccaca tacagatcca agaaacccgc taagaacctc
     1921 aagatgcccg gcgtctacta tgtggaccac agactggaaa gaatcaagga ggccgacaaa
     1981 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa
     2041 ctggggcaca aacttaattc cggactcaga tctcgagctc aagcttcgaa ttccaactga
     2101 aagctttgga attcgctagc cggccataca agtaatccgg atgatcgacg ccaacgtcgt
     2161 tgaattttca aattttaaat actgaatatt tgtttttttt cctattattt atttattctc
     2221 tttgtgtttt ttttcttgct ttctaaaaaa ttaattcaat ccaaatctaa acatgagggg
     2281 cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     2341 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2401 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2461 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3181 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3781 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
