LOCUS       sequence-492971-        3072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Lifeact-mNeonGreen (Gateway entry vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3072)
  TITLE     Flow Dynamics and CD44-HA Interactions Modulate Pancreatic Cancer 
            Cell attachment to endothelial cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3072)
  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            "Unpublished"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-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..3072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3..12
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      264..273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(289..339)
                     /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      336..345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(383..482)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(501..524)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(532..559)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(651..737)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(902..1490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(982..1001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1580..2389)
                     /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     2303..2322
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2381..2400
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    2512..2611
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(2629..3072,1..267))
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
ORIGIN
        1 taccatggcg gcctggaaag gcgacatccc gtcagggtag ggcaggtact gatggaagcc
       61 atacccgata tgagggacca gaatccaggg ggagaactgg aggtcaccct tggtggactt
      121 caggtttaac tcctcataac catcatttgg attgccggtg ccctgaccca ccatgtcaaa
      181 gtccacaccg ttgatggagc caaagatgtg taactcatgt gtcgctggga gagaggccat
      241 gttatcctcc tcgcccttgc tcaccatggt ggcgaccggt ggatccccct cctccttgct
      301 gatgctctcg aacttcttga tcaggtcggc cacgcccatg gtggcggtac cggatccagt
      361 cgactgaatt ggttccggcg ccagcctgct tttttgtaca aagttggcat tataaaaaag
      421 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
      481 tggggcccga gcttaagtaa ctaactaaca ggaagagttt gtagaaacgc aaaaaggcca
      541 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
      601 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
      661 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
      721 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcttagtagt tagacgtccc
      781 cgagatccat gctagcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1081 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1321 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1441 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1501 tgatcttttc tacggggtct gacgctcagt ggaacggggc ccaatctgaa taatgttaca
     1561 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     1621 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1681 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1741 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1801 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     1861 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1921 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     1981 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     2041 tttcacctga atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag
     2101 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     2161 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     2221 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg
     2281 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     2341 tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg
     2401 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     2461 caatgtaaca tcagagattt tgagacacgg gccagagctg cagctggatg gcaaataatg
     2521 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttctt
     2581 ataatgccaa ctttgtacaa gaaagctggg tctagatatc tcgagctatt acttgtacag
     2641 ctcgtccatg cccatcacat cggtaaaggc cttttgccac tccttgaagt tgagctcggt
     2701 cttggagtgc ttgagctccg tcttacggaa cacgtacatc ggctggttct tcagatagtt
     2761 agccgccatt ggcttggcaa aggtgtaggt ggtccgcgca gtgctccggt agcgcttgcc
     2821 atttccagtg gtgtaactcc acttaaaggt actgatgatg gttttgtcgt tggggtaagt
     2881 cttcttcgac ctgcaccagt ccgcagcggt cagcgagttg gtcatcacag gaccgtcagc
     2941 agggaaacca gtccccttca cctgggcctc tcctttgatg tggcttccct cgtaggtgta
     3001 gcggtagtta acagtaaggg aggcaccatc ttcaaactgc attgtgcgat ggacttggta
     3061 gccggagcca tc
//
