LOCUS       sequence-237327-        4448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Nuclear Localized mOrange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4448)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 4448)
  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::mOrange
FEATURES             Location/Qualifiers
     source          1..4448
                     /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="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2260..2280)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2260..2278)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2299..2315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2299..2315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2312..2334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2323..2339
                     /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(2347..2353,2354..2371,2372..2377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2392..2413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2530..2547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2701..3289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2781..2800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3460..4251,4252..4320))
                     /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     4083..4102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4321..4425)
                     /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 gtgagcaagg gcgaggagaa taacatggcc
     1381 atcatcaagg agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa cggccacgag
     1441 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gctttcagac cgctaagctg
     1501 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcatttcacc
     1561 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt caagctgtcc
     1621 ttccccgagg gcttcaagtg ggagcgcgtg atgaactacg aggacggcgg cgtggtgacc
     1681 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1741 accaacttcc cctccgacgg ccccgtgatg cagaagaaga ccatgggctg ggaggcctcc
     1801 tccgagcgga tgtaccccga ggacggtgcc ctgaagggca agatcaagat gaggctgaag
     1861 ctgaaggacg gcggccacta cacctccgag gtcaagacca cctacaaggc caagaagccc
     1921 gtgcagctgc ccggcgccta catcgtcgac atcaagttgg acatcacctc ccacaacgag
     1981 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2041 gacgagctgt acaagtaaaa gctttggaat tcgctagccg gccatacaag taatccggat
     2101 gatcgacgcc aacgtcgttg aattttcaaa ttttaaatac tgaatatttg ttttttttcc
     2161 tattatttat ttattctctt tgtgtttttt ttcttgcttt ctaaaaaatt aattcaatcc
     2221 aaatctaaac atgaggggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     2281 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2341 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2401 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2461 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2881 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3121 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4441 agtgccac
//
