LOCUS       sequence-443896-        6589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent probe for S/G2 cell cycle transition and G2/M 
            transition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6589)
  AUTHORS   Bajar BT, Lam AJ, Badiee RK, Oh YH, Chu J, Zhou XX, Kim N, Kim BB, 
            Chung M, Yablonovitch AL, Cruz BF, Kulalert K, Tao JJ, Meyer T, Su 
            XD, Lin MZ
  TITLE     Fluorescent indicators for simultaneous reporting of all four cell 
            cycle phases.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045.
  PUBMED    27798610
REFERENCE   2  (bases 1 to 6589)
  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: "Nat 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045."
COMMENT     Sequence Label: pLL3.7m-mTurquoise2-SLBP(18-126)-IRES-H1-mMaroon1
FEATURES             Location/Qualifiers
     source          1..6589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        129..432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(628..696,697..1488)
                     /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     complement(846..865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1659..2247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2148..2167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             2309..2489
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     2711..2727
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2712..2728
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2731..3319
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2837..2848
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3246..3266
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3375..3408
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4776..5348
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4935..4954)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5164..5181
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(5762..6472,6473..6475,6476..6478))
                     /codon_start=1
                     /product="monomeric optimized variant of ECFP (Kremers et 
                     al., 2006)"
                     /label=SCFP3A
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5804..5825)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6153..6172
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6412..6433
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(join(6514..6589,1..128))
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6543..6563)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
       61 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
      121 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
      181 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
      241 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
      301 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
      361 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
      421 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
      481 tattaataac tagtgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      601 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      661 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      721 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      781 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      841 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      901 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
      961 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1021 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1081 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1141 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1201 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1261 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1321 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1381 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1441 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1501 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1561 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1621 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1681 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1741 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1801 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1861 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1921 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1981 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2041 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2101 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2161 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2221 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2281 gccttttgct ggccttttgc tcacatgctg ctagagattt tccacactga ctaaaagggt
     2341 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     2401 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     2461 gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagtgg gagtgaatta
     2521 gcccttccag tccccccttt tcttttaaaa tcacgcgtca gctgccttgt aagtcattgg
     2581 tcttaaaggt acctgaggtg tgactggaaa acccacctcc tcctcctctt gtgcttctag
     2641 ccaggcacaa tcagcattgg tagctgctgt attgctactt gtgattgctc catgtttttc
     2701 taggtctcga tcgaggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
     2761 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
     2821 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
     2881 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
     2941 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     3001 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
     3061 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
     3121 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
     3181 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     3241 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     3301 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttgtacctag tggaaccgga
     3361 acccttaaac atgtataact tcgtataatg tatgctatac gaagttatta ggtccctcga
     3421 cgaattcgtt taaactcaat taagtttgtg ccccagtttg ctagggaggt cgcagtatct
     3481 ggccacagcc acctcgtgct gctcgacgta ggtctcattg tcggcctcct tgattctttc
     3541 cagtctcctg tcaataaaat agacgccggg catcttgagg ttctttgcgg gtttcttgga
     3601 tctgtatgtg gtttccagtc tgcagtgcag gtggcccccg cccacgagct tcagggccca
     3661 gtccagtctg ccttccaggc tgccgtcagc ggggtacagg gtctcggtac tggcctccca
     3721 gccgagtgtt ttcttctgca tcacagggcc gttggatggg aagttctccc ctctgacttg
     3781 gacgttgtag atcaggcagc cgtcctggag gctggtgtcc tgtgtagcag tgagtgtgcc
     3841 cccgtcttcg taaacggtgg ttctctccca tgtgaagccc tcagggaagg actgcttaaa
     3901 ataatcggga atgtcaggtg gatggttaat gaaggtcttg ctcccgtaca taaagcaggg
     3961 ggccaggatg tcgaatgcga acggcagggg gcctccgcgc accaccttga tcctgttggt
     4021 ctgggtgccc tcgtagggct tgccctcccc ttcggcggtg cattcgaagt agtggttgtt
     4081 cacggtgcct gtcaggtaca gcttggtgtg catgttctcc ttgatcagct cctcgccctt
     4141 gctcaccatg cctccggagc ctccagatct gagtccggac ttcttcttgc tccccctctt
     4201 ggcactcgac ttggctttgg gcttcacggt tttggccttc ttgggtttgg aggccttgac
     4261 tggtttgact ttgacaacct tgggcttttt ggctttcttg ggcgtggcag ccggcttctt
     4321 cttggccttc ttgacagggg tggctttggg tttcttgctt ggggctttgg aggcagcctt
     4381 cttgggcttg gctgccttct ttggagtggc cactttcttg acttccttct tggtcttctt
     4441 gaaagccacc gaccttttgg gctcatcgcc cttggccagc ctgaaggacc ccgaggcgcc
     4501 cacccctttg gtttgcttga gaacaccggt ggtcactagg cgcttgatgg acaacttgat
     4561 ctgggagtcg gcgttctcac ccaccttgta gtggctcttg atatactttt ggatggactg
     4621 gcgcgaggag ccggcacggt tcttctctgc ctggatagca gccacgatca tgtctgaata
     4681 cttggggtgg tccgtggact tcttggaagc cttggcccgt ttgggcttcg ccgccggggc
     4741 ggaggtggag ttctcggtca tgaattctgt ggccattatc atcgtgtttt tcaaaggaaa
     4801 accacgtccc cgtggttcgg ggggcctaga cgttttttaa cctcgactaa acacatgtaa
     4861 agcatgtgca ccgaggcccc agatcagatc ccatacaatg gggtaccttc tgggcatcct
     4921 tcagcccctt gttgaatacg cttgaggaga gccatttgac tctttccaca actatccaac
     4981 tcacaacgtg gcactggggt tgtgccgcct ttgcaggtgt atcttataca cgtggctttt
     5041 ggccgcagag gcacctgtcg ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag
     5101 acgttgtttg tcttcaagaa gcttccagag gaactgcttc cttcacgaca ttcaacagac
     5161 cttgcattcc tttggcgaga ggggaaagac ccctaggaat gctcgtcaag aagacagggc
     5221 caggtttccg ggccctcaca ttgccaaaag acggcaatat ggtggaaaat aacatataga
     5281 caaacgcaca ccggccttat tccaagcggc ttcggccagt aacgttaggg gggggggagg
     5341 gagaggggcc ttaagctaag atccttcttc atcctcgatc gtcagccggc acagtagaca
     5401 tagactcctt tgaatcagaa cttcctgatg atgattttct ctctcttcca aagtcattga
     5461 tgaggagttt ccttttatat cttgccattt ctttgttaac tctggtcctc atttcatctt
     5521 cttcaactgc acttgcccag tcagagcatc tggaacgggg tttagggcct tcaggagtgg
     5581 taaagctctc gggtctgcgc tcggcgccgc ggtgctctgc ctcctcggcg tcttcgggcc
     5641 tccagcgcct gccgtcggct ctgcgcttcc gtcccaggct ccatcgcgcg ggggacggcg
     5701 ggctggcctc gagtggtcgc cagtgtgatg ggtatcctcc cagggaaggc aacattgagt
     5761 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     5821 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt
     5881 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     5941 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     6001 tcttctgctt gtcggcggtg atatagacgt tgtcgctgaa gtagttgtac tccagcttgt
     6061 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     6121 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     6181 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     6241 tcatgtggtc ggggtagcgg gcgaagcact gcacgcccca gctcagggtg gtcacgaggg
     6301 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     6361 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     6421 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatac
     6481 ctggtgctag cggatctgac ggttcactaa accagctctg cttatataga cctcccaccg
     6541 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacat
//
