pUBC-mScarletI-1POK

LOCUS       pUBC-mScarletI-1POK               6330 bp DNA     circular SYN 12-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  AUTHORS   Ruizhao Wang
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..102
                     /label=mScarletI-1P0K-F
     primer_bind     73..99
                     /label=mScarletI-FLAG-Qch-F
     primer_bind     complement(73..99)
                     /label=mScarletI-FLAG-Qch-R
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             83..778
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(216..234)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     484..503
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(755..802)
                     /label=mCherry-Flag-R
     primer_bind     779..822
                     /label=Flag-1POK-F
     CDS             779..802
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             803..838
                     /codon_start=1
                     /label=Linker12
                     /translation="GGSGGTGGSGGT"
     CDS             839..2005
                     /codon_start=1
                     /label=1P0K(E239Y)
                     /translation="IDYTAAGFTLLQGAHLYAPEDRGICDVLVANGKIIAVASNIPSDI
                     VPNCTVVDLSGQILCPGFIDQHVHLIGGGGEAGPTTRTPEVALSRLTEAGVTSVVGLLG
                     TDSISRHPESLLAKTRALNEEGISAWMLTGAYHVPSRTITGSVEKDVAIIDRVIGVKCA
                     ISDHRSAAPDVYHLANMAAESRVGGLLGGKPGVTVFHMGDSKKALQPIYDLLENCDVPI
                     SKLLPTHVNRNVPLFYQALEFARKGGTIDITSSIDEPVAPAEGIARAVQAGIPLARVTL
                     SSDGNGSQPFFDDEGNLTHIGVAGFETLLETVQVLVKDYDFSISDALRPLTSSVAGFLN
                     LTGKGEILPGNDADLLVMTPELRIEQVYARGKLMVKDGKACVKGTFETA"
     primer_bind     1866..1875
                     /label=mScarletI-FLAG-Qch-F
     primer_bind     complement(1984..2029)
                     /label=1POK-NotI-R
     polyA_signal    2159..2214
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2575..2591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2599..2615
                     /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(2623..2653)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     2628..2649
                     /label=lac-pro-F
     protein_bind    2668..2689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2748..2943
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2773..2943
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     primer_bind     2789..2801
                     /label=mCherry-Flag-R
     rep_origin      2794..2929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2949..3083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3322..3910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3326..3345)
                     /label=pUBC-primer-1
     CDS             complement(4081..4941)
                     /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(4112..4133)
                     /label=Amp-F
     primer_bind     4855..4875
                     /label=Amp-R-seq
     promoter        complement(4942..5045)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5118..5517
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5245..5267)
                     /label=hUBC-pro-R
     primer_bind     5713..5736
                     /label=pUBC-seq-2
     promoter        5804..6330
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6167..6186
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gaagcttggg ctgcaggtcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
       61 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat
      121 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga
      181 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
      241 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt
      301 catcaagcac cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa
      361 gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc
      421 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga
      481 cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc
      541 cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg
      601 ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc
      661 ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga
      721 acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagga
      781 ttacaaggac gacgatgaca agggcggcag cggcggcacc ggcggcagcg gcggcaccat
      841 cgactacacc gccgccggct tcaccctgct gcagggcgcc cacctgtacg cccccgagga
      901 caggggcatc tgcgacgtgc tggtggccaa cggcaagatc atcgccgtgg ccagcaacat
      961 ccccagcgac atcgtgccca actgcaccgt ggtggacctg agcggccaga tcctgtgccc
     1021 cggcttcatc gaccagcacg tgcacctgat cggcggcggc ggcgaggccg gccccaccac
     1081 caggaccccc gaggtggccc tgagcaggct gaccgaggcc ggcgtgacca gcgtggtggg
     1141 cctgctgggc accgacagca tcagcaggca ccccgagagc ctgctggcca agaccagggc
     1201 cctgaacgag gagggcatca gcgcctggat gctgaccggc gcctaccacg tgcccagcag
     1261 gaccatcacc ggcagcgtgg agaaggacgt ggccatcatc gacagggtga tcggcgtgaa
     1321 gtgcgccatc agcgaccaca ggagcgccgc ccccgacgtg taccacctgg ccaacatggc
     1381 cgccgagagc agggtgggcg gcctgctggg cggcaagccc ggcgtgaccg tgttccacat
     1441 gggcgacagc aagaaggccc tgcagcccat ctacgacctg ctggagaact gcgacgtgcc
     1501 catcagcaag ctgctgccca cccacgtgaa caggaacgtg cccctgttct accaggccct
     1561 ggagttcgcc aggaagggcg gcaccatcga catcaccagc agcatcgacg agcccgtggc
     1621 ccccgccgag ggcatcgcca gggccgtgca ggccggcatc cccctggcca gggtgaccct
     1681 gagcagcgac ggcaacggca gccagccctt cttcgacgac gagggcaacc tgacccacat
     1741 cggcgtggcc ggcttcgaga ccctgctgga gaccgtgcag gtgctggtga aggactacga
     1801 cttcagcatc agcgacgccc tgaggcccct gaccagcagc gtggccggct tcctgaacct
     1861 gaccggcaag ggcgagatcc tgcccggcaa cgacgccgac ctgctggtga tgacccccga
     1921 gctgaggatc gagcaggtgt acgccagggg caagctgatg gtgaaggacg gcaaggcctg
     1981 cgtgaagggc accttcgaga cggcctgagc ggccgcactc ctcaggtgca ggctgcctat
     2041 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     2101 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     2161 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     2221 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     2281 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     2341 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     2401 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     2461 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     2521 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     2581 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2641 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2701 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     2761 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2881 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2941 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3001 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3061 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     3121 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3841 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3901 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4081 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4141 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4201 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4261 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4321 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4381 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4441 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4501 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4561 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4621 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4681 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4741 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4801 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4861 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4921 acggaaatgt tgaatactca tactcttcct ttttcatatt attgaagcat ttatcagggt
     4981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5041 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga ctagtaaccc
     5101 gtgtcggctc cagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     5161 cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc
     5221 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     5281 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     5341 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     5401 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     5461 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     5521 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     5581 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     5641 gggggttggg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     5701 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     5761 ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     5821 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
     5881 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
     5941 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
     6001 cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
     6061 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
     6121 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
     6181 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
     6241 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa
     6301 ttctggccgt ttttggcttt tttgttagac
//