LOCUS       sequence-237334-        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"
     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..2082
                     /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(2284..2304)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2284..2302)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2323..2339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2323..2339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2336..2358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2347..2363
                     /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(2371..2377,2378..2395,2396..2401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2416..2437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2554..2571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2725..3313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2805..2824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3484..4275,4276..4344))
                     /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     4107..4126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4345..4449)
                     /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 gcgaggagct gatcaaggag
     1381 aacatgcaca ccaagctgta catggaaggc accgtgaaca accaccactt caagtgcacc
     1441 cacgaagggg agggcaagcc ctacgagggc acccagacca acaggattaa ggtggtggag
     1501 ggaggccccc tgccgttcgc attcgacatc ctggccacct gctttatgta cgggagcaag
     1561 accttcatca accacaccca gggcatcccc gatttcttta agcagtcctt ccctgagggc
     1621 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgcttaccgt tacccaggac
     1681 accagcctcc aggacggctg cttgatctac aacgtcaagc tcagaggggt gaacttccca
     1741 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccagtac cgagaccctg
     1801 taccccgctg acggcggcct ggaaggcaga tgcgacatgg ccctgaagct cgtgggcggg
     1861 ggccacctgc actgcaacct gaagaccaca tacagatcca agaaacccgc taagaacctc
     1921 aagatgcccg gcgtctactt tgtggaccgc agactggaaa gaatcaagga ggccgacaat
     1981 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa
     2041 ctggggcaca aacttaatgg catggacgag ctgtacaagt aaaagctttg gaattcgcta
     2101 gccggccata caagtaatcc ggatgatcga cgccaacgtc gttgaatttt caaattttaa
     2161 atactgaata tttgtttttt ttcctattat ttatttattc tctttgtgtt ttttttcttg
     2221 ctttctaaaa aattaattca atccaaatct aaacatgagg ggcccggtac ccagcttttg
     2281 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2341 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2461 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4441 ggttccgcgc acatttcccc gaaaagtgcc ac
//
