LOCUS       sequence-241714-        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"
     primer_bind     37..59
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     51..68
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     52..68
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     78..97
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        78..96
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             800..1510
                     /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(1712..1732)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1712..1730)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1751..1767)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1751..1767)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1764..1786)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1775..1791
                     /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(1799..1805,1806..1823,1824..1829))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1844..1865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1982..1999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2153..2741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2233..2252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2912..3703,3704..3772))
                     /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     3535..3554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3773..3877)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3903..4358)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4040..4061)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
       61 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggac
      121 tagtggatcc cccgggctgc agcctaggat gtctgattcc gctgttgttg ccgccgctgt
      181 cgagccaaag gtcccaaagg ctaaggccgc caaggccgcc aagccagccg ccaaggccgc
      241 caagccaacc aaagttgcca aggccaaggc tccagtcgct cacccaccat acatcaacat
      301 gatcaaggag gccatcaagc agctcaagga ccgcaaagga gcttccaagc aggcaatcct
      361 caagttcatc tcccaaaact acaagctcgg agacaatgtc atccagatca atgctcatct
      421 ccgtcaggct ctcaagcgtg gtgtcaccag caaggccctt gttcaagccg ccggatccgg
      481 agccaacgga cgtttccgtg tgccagagaa ggccgccgcc gccaagaagc cagcagcagc
      541 caagaagcca gcagcagcca agaaaccagc agccgccaag aaagccactg gagagaagaa
      601 ggcaaagaag ccagccgctg ccaagccaaa gaaggccgcc accggagata agaaggtcaa
      661 gaaggccaag agcccaaaga aggttgccaa gccagctgcc aagaaggtcg ccaagtctcc
      721 agccaagaag gccgctccaa agaagatcgc caagccagct gccaagaagg ccgccaagcc
      781 agccgccaag gccgcatgca tggtgagcaa gggcgaggag aataacatgg ccatcatcaa
      841 ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat
      901 cgagggcgag ggcgagggcc gcccctacga gggctttcag accgctaagc tgaaggtgac
      961 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcatttca cctacggctc
     1021 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttcaagctgt ccttccccga
     1081 gggcttcaag tgggagcgcg tgatgaacta cgaggacggc ggcgtggtga ccgtgaccca
     1141 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1201 cccctccgac ggccccgtga tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1261 gatgtacccc gaggacggtg ccctgaaggg caagatcaag atgaggctga agctgaagga
     1321 cggcggccac tacacctccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1381 gcccggcgcc tacatcgtcg acatcaagtt ggacatcacc tcccacaacg aggactacac
     1441 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1501 gtacaagtaa aagctttgga attcgctagc cggccataca agtaatccgg atgatcgacg
     1561 ccaacgtcgt tgaattttca aattttaaat actgaatatt tgtttttttt cctattattt
     1621 atttattctc tttgtgtttt ttttcttgct ttctaaaaaa ttaattcaat ccaaatctaa
     1681 acatgagggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     1741 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1801 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1861 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1921 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1981 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2041 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2101 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2161 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2221 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2281 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2341 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2401 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2461 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2521 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2581 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2641 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2701 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2761 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2821 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2881 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2941 tctcagcgat ctgcctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3001 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3061 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3181 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3841 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3901 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3961 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4021 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4081 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4141 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4201 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aatggaagaa
     4261 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4321 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     4381 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4441 gggatgtg
//
