LOCUS       sequence-241712-        5012 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 5012)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 5012)
  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::mOrange
FEATURES             Location/Qualifiers
     source          1..5012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..79
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     71..88
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     72..88
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     98..117
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        98..116
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             820..1530
                     /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(2296..2316)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2296..2314)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2335..2351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2335..2351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2348..2370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2359..2375
                     /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(2383..2389,2390..2407,2408..2413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2428..2449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2566..2583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2737..3325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2817..2836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3496..4287,4288..4356))
                     /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     4119..4138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4357..4461)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4487..4942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4624..4645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4836..4855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
       61 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
      121 attggagctc caccgcggac tagtggatcc cccgggctgc agcctaggat gtctgattcc
      181 gctgttgttg ccgccgctgt cgagccaaag gtcccaaagg ctaaggccgc caaggccgcc
      241 aagccagccg ccaaggccgc caagccaacc aaagttgcca aggccaaggc tccagtcgct
      301 cacccaccat acatcaacat gatcaaggag gccatcaagc agctcaagga ccgcaaagga
      361 gcttccaagc aggcaatcct caagttcatc tcccaaaact acaagctcgg agacaatgtc
      421 atccagatca atgctcatct ccgtcaggct ctcaagcgtg gtgtcaccag caaggccctt
      481 gttcaagccg ccggatccgg agccaacgga cgtttccgtg tgccagagaa ggccgccgcc
      541 gccaagaagc cagcagcagc caagaagcca gcagcagcca agaaaccagc agccgccaag
      601 aaagccactg gagagaagaa ggcaaagaag ccagccgctg ccaagccaaa gaaggccgcc
      661 accggagata agaaggtcaa gaaggccaag agcccaaaga aggttgccaa gccagctgcc
      721 aagaaggtcg ccaagtctcc agccaagaag gccgctccaa agaagatcgc caagccagct
      781 gccaagaagg ccgccaagcc agccgccaag gccgcatgca tggtgagcaa gggcgaggag
      841 aataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg
      901 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggctttcag
      961 accgctaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1021 cctcatttca cctacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     1081 ttcaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaacta cgaggacggc
     1141 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1201 aagctgcgcg gcaccaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc
     1261 tgggaggcct cctccgagcg gatgtacccc gaggacggtg ccctgaaggg caagatcaag
     1321 atgaggctga agctgaagga cggcggccac tacacctccg aggtcaagac cacctacaag
     1381 gccaagaagc ccgtgcagct gcccggcgcc tacatcgtcg acatcaagtt ggacatcacc
     1441 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     1501 accggcggca tggacgagct gtacaagtaa aagctttgga attcatctcg cgcccgtgcc
     1561 tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc
     1621 caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct
     1681 tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa
     1741 aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac
     1801 acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca
     1861 tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat
     1921 ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag
     1981 ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc
     2041 ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg
     2101 ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg
     2161 aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt
     2221 ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt tgggcccggt
     2281 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2341 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag
     2401 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2461 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2521 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2581 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2641 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2701 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2761 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2821 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2881 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2941 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3001 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3061 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3121 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3181 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgcct
     3541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3781 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4081 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     4501 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4561 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4621 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4681 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4741 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4801 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4861 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4921 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4981 cgatcggtgc gggcctcttc gctattacgc ca
//
