LOCUS       sequence-382201-        7936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of mTurquoise in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7936)
  AUTHORS   Silverio-Alves R, Kurochkin I, Rydstrom A, Vazquez Echegaray C, 
            Haider J, Nicholls M, Rode C, Thelaus L, Lindgren AY, Ferreira AG, 
            Brandao R, Larsson J, de Bruijn MFTR, Martin-Gonzalez J, Pereira CF
  TITLE     GATA2 mitotic bookmarking is required for definitive haematopoiesis.
  JOURNAL   Nat Commun. 2023 Aug 14;14(1):4645. doi: 10.1038/s41467-023-40391-x.
  PUBMED    37580379
REFERENCE   2  (bases 1 to 7936)
  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 
            Commun."; date: "2023-08-14"; volume: "14(1):4645. doi"; pages: " 
            10.1038/s41467-023-40391-x"
COMMENT     Sequence Label: FUW-mTurquoise
FEATURES             Location/Qualifiers
     source          1..7936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..55)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        227..606
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        608..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     761..781
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             824..1004
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1051..1176
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1669..1902
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2087..2131
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2429..2546
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2611..3822
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3659..3678
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(3843..3845,3846..3848,3849..4562)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3888..3909)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4149..4168)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4496..4517
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4591..5179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4644..4664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5062..5073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5182..5198)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5183..5199)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5704..5884
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5910..5927)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5916..6027
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6173..6761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6253..6272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6932..7723,7724..7792))
                     /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     7555..7574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7793..7897)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
       61 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
      121 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      181 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      241 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      301 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      361 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      421 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      481 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      541 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      601 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      661 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      721 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      781 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
      841 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      901 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      961 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     1021 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     1081 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     1141 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     1201 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1261 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1321 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1381 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1441 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1501 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     1561 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     1621 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     1681 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     1741 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     1801 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     1861 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     1921 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     1981 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     2041 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     2101 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     2161 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     2221 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     2281 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     2341 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
     2401 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     2461 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     2521 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     2581 ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg gcgcctcccg
     2641 cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc
     2701 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
     2761 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
     2821 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
     2881 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
     2941 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
     3001 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
     3061 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
     3121 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     3181 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     3241 ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg
     3301 agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     3361 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3421 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3481 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3541 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     3601 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     3661 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     3721 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa
     3781 ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagcttg ggcccgggat
     3841 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3901 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3961 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4021 ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
     4081 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4141 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4201 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4261 acaagctgga gtacaactac atcagcgaca acgtctatat caccgccgac aagcagaaga
     4321 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     4381 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4441 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     4501 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     4561 aaggatccat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     4621 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4681 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4741 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4801 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4861 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4921 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4981 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     5041 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5101 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5161 ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca
     5221 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     5281 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     5341 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5401 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     5461 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
     5521 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
     5581 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
     5641 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt
     5701 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5761 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5821 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5881 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     5941 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6001 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6061 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgtgag
     6121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7921 ctgacgtcga cggatc
//
