LOCUS       sequence-83989-e        4950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains membrane localized cerulean fluorescent protein in pCS 
            expression vector. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4950)
  AUTHORS   Megason SG
  TITLE     In toto imaging of embryogenesis with confocal time-lapse 
            microscopy.
  JOURNAL   Methods Mol Biol. 2009;546:317-32. doi: 
            10.1007/978-1-60327-977-2_19.
  PUBMED    19378112
REFERENCE   2  (bases 1 to 4950)
  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"; journalName: "Methods Mol
            Biol."; date: "2009"; volume: "546:317-32. doi"; pages: " 
            10.1007/978-1-60327-977-2_19"
COMMENT     Sequence Label: pCS-membrane-ceruleanFP
FEATURES             Location/Qualifiers
     source          1..4950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    4..138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(28..47)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     82..101
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(251..271)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(251..269)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(290..306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(290..306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(303..325)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    314..330
                     /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(338..344,345..362,363..368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    383..404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(521..538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(692..1280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(772..791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1451..2242,2243..2311))
                     /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     2074..2093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2312..2416)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2442..2897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2579..2600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2791..2810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2971..3955
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3990..4008
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3990..4007
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      4076..4085
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4130..4139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4193..4202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4199..4201,4202..4204,4205..4918)
                     /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(4244..4265)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4505..4524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4852..4873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
       61 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      121 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
      181 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
      241 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
      301 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      361 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      421 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      481 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      541 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      601 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      661 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      721 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      781 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      841 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      901 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      961 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1021 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1081 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     1141 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1201 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1261 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1321 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1381 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1441 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1501 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1561 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     1621 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1681 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1741 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1801 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1861 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1921 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1981 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2041 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2101 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2161 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2221 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2281 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2341 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2401 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     2461 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2521 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2581 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2641 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2701 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2761 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2821 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2881 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2941 ggtgcgggcc tcttcgctat tacgccagtc gaccatagcc aattcaatat ggcgtatatg
     3001 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
     3061 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     3121 gcaccatgcc aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt
     3181 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     3241 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     3301 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     3361 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     3421 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     3481 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     3541 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     3601 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     3661 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     3721 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca
     3781 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     3841 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     3901 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaaccgcca
     3961 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
     4021 tgttcttttt gcaggatccc atcgattcga attcaaggcc tctcgagcct ctagagccac
     4081 catgggctgc atcaagagca agcgcaagga caacctgaac gacgacgagg ccgccatggg
     4141 ctgcatcaag agcaagcgca aggacaacct gaacgacgac gaggcaccgg tcgccaccat
     4201 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4261 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4321 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4381 cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     4441 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4501 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4561 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4621 gctggagtac aacgccatca gcgacaacgt ctatatcacc gccgacaagc agaagaacgg
     4681 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4741 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4801 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4861 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     4921 cggccctaga actatagtga gtcgtattac
//
