LOCUS       sequence-241693-        5021 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 5021)
  TITLE     Leifer Lab worm expression plasmids
REFERENCE   2  (bases 1 to 5021)
  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::Cerulean
FEATURES             Location/Qualifiers
     source          1..5021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..60
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     52..69
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     53..69
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     79..98
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        79..97
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(801..803,804..806,807..1520)
                     /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(846..867)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1107..1126)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1454..1475
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2286..2306)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2286..2304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2325..2341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2325..2341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2338..2360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2349..2365
                     /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(2373..2379,2380..2397,2398..2403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2418..2439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2556..2573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2727..3315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2807..2826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3486..4277,4278..4346))
                     /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     4109..4128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4347..4451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4477..4932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4614..4635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4826..4845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
       61 acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct ccaccgcgga
      121 ctagtggatc ccccgggctg cagcctagga tgtctgattc cgctgttgtt gccgccgctg
      181 tcgagccaaa ggtcccaaag gctaaggccg ccaaggccgc caagccagcc gccaaggccg
      241 ccaagccaac caaagttgcc aaggccaagg ctccagtcgc tcacccacca tacatcaaca
      301 tgatcaagga ggccatcaag cagctcaagg accgcaaagg agcttccaag caggcaatcc
      361 tcaagttcat ctcccaaaac tacaagctcg gagacaatgt catccagatc aatgctcatc
      421 tccgtcaggc tctcaagcgt ggtgtcacca gcaaggccct tgttcaagcc gccggatccg
      481 gagccaacgg acgtttccgt gtgccagaga aggccgccgc cgccaagaag ccagcagcag
      541 ccaagaagcc agcagcagcc aagaaaccag cagccgccaa gaaagccact ggagagaaga
      601 aggcaaagaa gccagccgct gccaagccaa agaaggccgc caccggagat aagaaggtca
      661 agaaggccaa gagcccaaag aaggttgcca agccagctgc caagaaggtc gccaagtctc
      721 cagccaagaa ggccgctcca aagaagatcg ccaagccagc tgccaagaag gccgccaagc
      781 cagccgccaa ggccgcatgc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      841 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      901 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      961 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc
     1021 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1081 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1141 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1201 acggcaacat cctggggcac aagctggagt acaacgccat cagcgacaac gtctatatca
     1261 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1321 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1381 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     1441 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1501 tggacgagct gtacaagtaa aagctttgga attcatctcg cgcccgtgcc tctgacttct
     1561 aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat
     1621 ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac
     1681 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     1741 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     1801 gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac
     1861 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     1921 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     1981 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     2041 tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac
     2101 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     2161 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     2221 aatgacagaa tacattccca atataccaaa cataactgtt tgggcccggt acccagcttt
     2281 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     2341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     2401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgcct atttcgttca
     3541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4441 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     4501 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4561 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4621 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4681 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4741 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4801 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4861 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4921 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4981 gggcctcttc gctattacgc cagctggcga aagggggatg t
//
