LOCUS       sequence-241915-        5036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Nuclear Localized mNeptune.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5036)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 5036)
  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::mNeptunr
FEATURES             Location/Qualifiers
     source          1..5036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..87
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     79..96
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     80..96
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     106..125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        106..124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             828..1562
                     /codon_start=1
                     /product="brighter version of the far-red fluorescent 
                     protein mNeptune2 with slightly blue-shifted spectra"
                     /label=mNeptune2.5
                     /translation="MVSKGEELIKENMHTKLYMEGTVNNHHFKCTHEGEGKPYEGTQTN
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(2328..2348)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2328..2346)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2367..2383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2367..2383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2380..2402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2391..2407
                     /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(2415..2421,2422..2439,2440..2445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2460..2481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2598..2615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2769..3357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2849..2868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3528..4319,4320..4388))
                     /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     4151..4170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4389..4493)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4519..4974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4656..4677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4868..4887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
       61 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
      121 tagggcgaat tggagctcca ccgcggacta gtggatcccc cgggctgcag cctaggatgt
      181 ctgattccgc tgttgttgcc gccgctgtcg agccaaaggt cccaaaggct aaggccgcca
      241 aggccgccaa gccagccgcc aaggccgcca agccaaccaa agttgccaag gccaaggctc
      301 cagtcgctca cccaccatac atcaacatga tcaaggaggc catcaagcag ctcaaggacc
      361 gcaaaggagc ttccaagcag gcaatcctca agttcatctc ccaaaactac aagctcggag
      421 acaatgtcat ccagatcaat gctcatctcc gtcaggctct caagcgtggt gtcaccagca
      481 aggcccttgt tcaagccgcc ggatccggag ccaacggacg tttccgtgtg ccagagaagg
      541 ccgccgccgc caagaagcca gcagcagcca agaagccagc agcagccaag aaaccagcag
      601 ccgccaagaa agccactgga gagaagaagg caaagaagcc agccgctgcc aagccaaaga
      661 aggccgccac cggagataag aaggtcaaga aggccaagag cccaaagaag gttgccaagc
      721 cagctgccaa gaaggtcgcc aagtctccag ccaagaaggc cgctccaaag aagatcgcca
      781 agccagctgc caagaaggcc gccaagccag ccgccaaggc cgcatgcatg gtgagcaagg
      841 gcgaggagct gatcaaggag aacatgcaca cgaagctgta catggaaggc accgtgaaca
      901 accaccactt caagtgcacc cacgaagggg agggcaagcc ctacgagggc acccagacca
      961 acaggattaa ggtggtggag ggaggccccc tgccgttcgc attcgacatc ctggccacct
     1021 gctttatgta cgggagcaag accttcatca accacaccca gggcatcccc gatttcttta
     1081 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     1141 tgcttaccgt tacccaggac accagcctcc aggacggctg cttgatctac aacgtcaagc
     1201 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     1261 aggccagtac cgagaccctg taccccgctg acggcggcct ggaaggcaga tgcgacatgg
     1321 ccctgaagct cgtgggcggg ggccacctgc actgcaacct gaagaccaca tacagatcca
     1381 agaaacccgc taagaacctc aagatgcccg gcgtctactt tgtggaccgc agactggaaa
     1441 gaatcaagga ggccgacaat gagacctacg tcgagcagca cgaggtggct gtggccagat
     1501 actgcgacct ccctagcaaa ctggggcaca aacttaatgg catggacgag ctgtacaagt
     1561 aaaagctttg gaattcatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca
     1621 acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa
     1681 acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt
     1741 gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc
     1801 cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt
     1861 ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca aaatacctta
     1921 tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct
     1981 catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat
     2041 caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt cccctattgt
     2101 ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga
     2161 tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg
     2221 ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc
     2281 caatatacca aacataactg tttgggcccg gtacccagct tttgttccct ttagtgaggg
     2341 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2401 ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa
     2461 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2521 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2581 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2641 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2701 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2761 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2821 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2881 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2941 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3001 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3061 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3541 atcagtgagg cacctatctc agcgatctgc ctatttcgtt catccatagt tgcctgactc
     3601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4501 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4561 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4621 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4681 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4741 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4801 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4861 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4921 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4981 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgcta
//
