LOCUS       sequence-492972-        3060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Lifeact-mScarlet-I (Gateway entry vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3060)
  TITLE     Flow Dynamics and CD44-HA Interactions Modulate Pancreatic Cancer 
            Cell attachment to endothelial cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3060)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pENTR2b-Lifeact-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..3060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(390..1199)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1113..1132
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1191..1210
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1322..1421
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1442..2137)
                     /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(1717..1736)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1986..2004
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2134..2143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2159..2209)
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      2206..2215
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2253..2352)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2371..2394)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(2402..2429)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2521..2607)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(2772..3060,1..300))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2852..2871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
       61 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacggggc ccaatctgaa
      361 taatgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
      421 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
      481 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
      541 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
      601 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
      661 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
      721 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
      781 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
      841 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgtttttccg
      901 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
      961 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1021 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag
     1081 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1141 tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata
     1201 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     1261 ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccagagctg cagctggatg
     1321 gcaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca
     1381 atgctttctt ataatgccaa ctttgtacaa gaaagctggg tctagatatc tcgagctatt
     1441 acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggagcgt tcgtactgtt
     1501 ccaccacggt gtagtcctcg ttgtgggagg tgatgtccaa cttgcggtcg acgttgtagg
     1561 cgccgggcat ctgcacgggc ttcttggcct tgtaggtggt cttgaagtcc gccaggtagc
     1621 ggccgccgtc cttcaggcgc agggccatct taatgtcgcc cttcagcacg ccgtcctcgg
     1681 ggtacaaccg ctcggtggac gcttcccagc ccattgtctt cttctgcatt acggggccgt
     1741 caggagggaa gttggtgccg cggagcttca ccttgtagat cagggtgccg tcctccaggg
     1801 aggtgtcctg ggtcacggtc acggcgccgc cgtcctcgaa gttcatcacg cgctcccact
     1861 tgaagccctc ggggaaggac tgcttatagt agtcggggat gtcggcgggg tgcttgatga
     1921 aggccctgga gccgtacatg aactgagggg acaggatgtc ccaggagaag ggcagggggc
     1981 cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct
     2041 cgccctcgat ctcgaactcg tggccgttca tggagccctc catgtgcacc ttgaaccgca
     2101 tgaactcctt gatcactgcc tcgcccttgc tcaccatggt ggcgaccggt ggatccccct
     2161 cctccttgct gatgctctcg aacttcttga tcaggtcggc cacgcccatg gtggcggtac
     2221 cggatccagt cgactgaatt ggttccggcg ccagcctgct tttttgtaca aagttggcat
     2281 tataaaaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2341 aatcattatt tggggcccga gcttaagtaa ctaactaaca ggaagagttt gtagaaacgc
     2401 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     2461 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     2521 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     2581 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcttagtagt
     2641 tagacgtccc cgagatccat gctagcggta atacggttat ccacagaatc aggggataac
     2701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
//
