LOCUS       sequence-241910-        5054 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 5054)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 5054)
  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: pAS1-his-24::tagRFP
FEATURES             Location/Qualifiers
     source          1..5054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..92
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     84..101
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     85..101
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     111..130
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             833..1543
                     /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(2351..2371)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2351..2369)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2390..2406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2390..2406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2403..2425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2414..2430
                     /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(2438..2444,2445..2462,2463..2468))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2483..2504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2621..2638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2792..3380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2872..2891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3551..4342,4343..4411))
                     /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     4174..4193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4412..4516)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4542..4997)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4679..4700)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4891..4910
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
       61 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
      121 cactataggg cgaattggag ctccaccgcg gactagtgga tcccccgggc tgcagcctag
      181 gatgtctgat tccgctgttg ttgccgccgc tgtcgagcca aaggtcccaa aggctaaggc
      241 cgccaaggcc gccaagccag ccgccaaggc cgccaagcca accaaagttg ccaaggccaa
      301 ggctccagtc gctcacccac catacatcaa catgatcaag gaggccatca agcagctcaa
      361 ggaccgcaaa ggagcttcca agcaggcaat cctcaagttc atctcccaaa actacaagct
      421 cggagacaat gtcatccaga tcaatgctca tctccgtcag gctctcaagc gtggtgtcac
      481 cagcaaggcc cttgttcaag ccgccggatc cggagccaac ggacgtttcc gtgtgccaga
      541 gaaggccgcc gccgccaaga agccagcagc agccaagaag ccagcagcag ccaagaaacc
      601 agcagccgcc aagaaagcca ctggagagaa gaaggcaaag aagccagccg ctgccaagcc
      661 aaagaaggcc gccaccggag ataagaaggt caagaaggcc aagagcccaa agaaggttgc
      721 caagccagct gccaagaagg tcgccaagtc tccagccaag aaggccgctc caaagaagat
      781 cgccaagcca gctgccaaga aggccgccaa gccagccgcc aaggccgcat gcatggtgtc
      841 taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt
      901 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
      961 gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     1021 taccagcttc atgtacggca gcagaacctt catcaaccac acccagggca tccccgactt
     1081 ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg
     1141 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     1201 caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg
     1261 ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag gcagaaccga
     1321 catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga ccacatacag
     1381 atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg accacagact
     1441 ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc
     1501 cagatactgc gacctcccta gcaaactggg gcacaaactt aattccggac tcagatctcg
     1561 agctcaagct tcgaattcca actgaaagct ttggaattca tctcgcgccc gtgcctctga
     1621 cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt gctcccaccc
     1681 cctatttttg ttattatcaa aaaacttctc ttaatttctt tgttttttag cttcttttaa
     1741 gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt
     1801 cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat
     1861 gttctgtgta cacttcttat gttttttact tctgataaat ttttttgaaa catcatagaa
     1921 aaaaccgcac acaaaatacc ttatcatatg ttacgtttca gtttatgacc gcaattttta
     1981 tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg
     2041 gagtattttt ggaatttttc aatcaagtga aagtttatga aattaatttt cctgcttttg
     2101 ctttttgggg tttcccctat tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa
     2161 atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct
     2221 cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg
     2281 ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttgggc ccggtaccca
     2341 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2401 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
     2461 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2521 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2581 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2641 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2701 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2881 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgcctatttc
     3601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4561 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4621 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4681 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4741 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4801 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4861 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4921 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4981 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5041 ggtgcgggcc tctt
//
