LOCUS       sequence-241916-        4472 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 4472)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 4472)
  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::mNeptune
FEATURES             Location/Qualifiers
     source          1..4472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..48
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     40..57
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     41..57
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     67..86
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        67..85
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             789..1523
                     /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(1725..1745)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1725..1743)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1764..1780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1764..1780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1777..1799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1788..1804
                     /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(1812..1818,1819..1836,1837..1842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1857..1878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1995..2012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2166..2754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2246..2265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2925..3716,3717..3785))
                     /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     3548..3567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3786..3890)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3916..4371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4053..4074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4265..4284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
       61 cgcgcgtaat acgactcact atagggcgaa ttggagctcc accgcggact agtggatccc
      121 ccgggctgca gcctaggatg tctgattccg ctgttgttgc cgccgctgtc gagccaaagg
      181 tcccaaaggc taaggccgcc aaggccgcca agccagccgc caaggccgcc aagccaacca
      241 aagttgccaa ggccaaggct ccagtcgctc acccaccata catcaacatg atcaaggagg
      301 ccatcaagca gctcaaggac cgcaaaggag cttccaagca ggcaatcctc aagttcatct
      361 cccaaaacta caagctcgga gacaatgtca tccagatcaa tgctcatctc cgtcaggctc
      421 tcaagcgtgg tgtcaccagc aaggcccttg ttcaagccgc cggatccgga gccaacggac
      481 gtttccgtgt gccagagaag gccgccgccg ccaagaagcc agcagcagcc aagaagccag
      541 cagcagccaa gaaaccagca gccgccaaga aagccactgg agagaagaag gcaaagaagc
      601 cagccgctgc caagccaaag aaggccgcca ccggagataa gaaggtcaag aaggccaaga
      661 gcccaaagaa ggttgccaag ccagctgcca agaaggtcgc caagtctcca gccaagaagg
      721 ccgctccaaa gaagatcgcc aagccagctg ccaagaaggc cgccaagcca gccgccaagg
      781 ccgcatgcat ggtgagcaag ggcgaggagc tgatcaagga gaacatgcac acgaagctgt
      841 acatggaagg caccgtgaac aaccaccact tcaagtgcac ccacgaaggg gagggcaagc
      901 cctacgaggg cacccagacc aacaggatta aggtggtgga gggaggcccc ctgccgttcg
      961 cattcgacat cctggccacc tgctttatgt acgggagcaa gaccttcatc aaccacaccc
     1021 agggcatccc cgatttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
     1081 ccacatacga agacgggggc gtgcttaccg ttacccagga caccagcctc caggacggct
     1141 gcttgatcta caacgtcaag ctcagagggg tgaacttccc atccaacggc cctgtgatgc
     1201 agaagaaaac actcggctgg gaggccagta ccgagaccct gtaccccgct gacggcggcc
     1261 tggaaggcag atgcgacatg gccctgaagc tcgtgggcgg gggccacctg cactgcaacc
     1321 tgaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     1381 ttgtggaccg cagactggaa agaatcaagg aggccgacaa tgagacctac gtcgagcagc
     1441 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatg
     1501 gcatggacga gctgtacaag taaaagcttt ggaattcgct agccggccat acaagtaatc
     1561 cggatgatcg acgccaacgt cgttgaattt tcaaatttta aatactgaat atttgttttt
     1621 tttcctatta tttatttatt ctctttgtgt tttttttctt gctttctaaa aaattaattc
     1681 aatccaaatc taaacatgag gggcccggta cccagctttt gttcccttta gtgagggtta
     1741 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1801 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     1861 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1921 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1981 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2041 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2101 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2161 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2581 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2941 agtgaggcac ctatctcagc gatctgccta tttcgttcat ccatagttgc ctgactcccc
     3001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3901 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3961 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4021 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4081 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4141 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4201 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4261 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4321 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4381 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4441 agctggcgaa agggggatgt gctgcaaggc ga
//
