LOCUS       sequence-241700-        4457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Nuclear Localized Cerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4457)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 4457)
  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::Cerulean
FEATURES             Location/Qualifiers
     source          1..4457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..77
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     69..86
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     70..86
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     96..115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        96..114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(818..820,821..823,824..1537)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(863..884)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1124..1143)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1471..1492
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1739..1759)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1739..1757)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1778..1794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1778..1794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1791..1813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1802..1818
                     /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(1826..1832,1833..1850,1851..1856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1871..1892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2009..2026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2180..2768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2260..2279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2939..3730,3731..3799))
                     /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     3562..3581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3800..3904)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3930..4385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4067..4088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4279..4298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
       61 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
      121 tggagctcca ccgcggacta gtggatcccc cgggctgcag cctaggatgt ctgattccgc
      181 tgttgttgcc gccgctgtcg agccaaaggt cccaaaggct aaggccgcca aggccgccaa
      241 gccagccgcc aaggccgcca agccaaccaa agttgccaag gccaaggctc cagtcgctca
      301 cccaccatac atcaacatga tcaaggaggc catcaagcag ctcaaggacc gcaaaggagc
      361 ttccaagcag gcaatcctca agttcatctc ccaaaactac aagctcggag acaatgtcat
      421 ccagatcaat gctcatctcc gtcaggctct caagcgtggt gtcaccagca aggcccttgt
      481 tcaagccgcc ggatccggag ccaacggacg tttccgtgtg ccagagaagg ccgccgccgc
      541 caagaagcca gcagcagcca agaagccagc agcagccaag aaaccagcag ccgccaagaa
      601 agccactgga gagaagaagg caaagaagcc agccgctgcc aagccaaaga aggccgccac
      661 cggagataag aaggtcaaga aggccaagag cccaaagaag gttgccaagc cagctgccaa
      721 gaaggtcgcc aagtctccag ccaagaaggc cgctccaaag aagatcgcca agccagctgc
      781 caagaaggcc gccaagccag ccgccaaggc cgcatgcatg gtgagcaagg gcgaggagct
      841 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      901 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      961 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg
     1021 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1081 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1141 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1201 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acgccatcag
     1261 cgacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1321 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1381 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     1441 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1501 cgggatcact ctcggcatgg acgagctgta caagtaaaag ctttggaatt cgctagccgg
     1561 ccatacaagt aatccggatg atcgacgcca acgtcgttga attttcaaat tttaaatact
     1621 gaatatttgt tttttttcct attatttatt tattctcttt gtgttttttt tcttgctttc
     1681 taaaaaatta attcaatcca aatctaaaca tgaggggccc ggtacccagc ttttgttccc
     1741 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1801 attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct
     1861 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1921 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1981 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2941 accaatgctt aatcagtgag gcacctatct cagcgatctg cctatttcgt tcatccatag
     3001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3901 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     3961 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4021 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4081 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4141 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4201 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4261 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4321 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4381 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4441 ttcgctatta cgccagc
//
