LOCUS       sequence-249586-        4295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA Kit # 1000000119 extension.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4295)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4295)
  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"
COMMENT     Sequence Label: pRDAX-VALENTIA-027   LifeAct-mVenus//BsaI
FEATURES             Location/Qualifiers
     source          1..4295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        158..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..814
                     /label=chicken beta-actin promoter
     regulatory      834..843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             840..890
                     /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"
     primer_bind     complement(1212..1231)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1680..1801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1717..1736)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1771..1790
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      2602..2631
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2653..3240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3141..3160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3252..3279)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3303..4094,4095..4163))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3926..3945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4164..4266)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4266..4295,1..2))
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctttctgcta tggaggtcag gtatgattta aaggtctcat aggtcttgta ttcagtctcg
       61 cactttagac atggtcttct gctgcccgtg gtcctctatt cgtcggatgt tctgtcgatt
      121 cttcctgtct atctggactg gatctgatat catcgtcgac attgattatt gactagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
      541 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
      601 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggggcg
      661 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
      721 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      781 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgttgg gacgccacca
      841 tgggcgtggc cgacctgatc aagaagttcg agagcatcag caaggaggag ggcgggtcag
      901 gcggcagcgt gtcaaaggga gaagagttgt ttaccggtgt tgttcccata ctggtggaac
      961 tggatggtga cgtcaatgga cacaagttta gcgtttctgg cgaaggagaa ggcgatgcta
     1021 cctatggcaa actgaccctc aaactgatct gtacgactgg aaagttgcct gtaccttggc
     1081 caacactggt cacaaccttg ggctatgggc tgcaatgctt tgcccgttat cctgatcaca
     1141 tgaagcaaca tgacttcttt aagtctgcta tgccagaggg atatgtgcag gaacggacta
     1201 tcttcttcaa ggacgacggc aactacaaga cgagagctga ggtgaaattt gagggtgaca
     1261 cactggtgaa tcgcatagag ctgaaaggca tcgacttcaa agaagatggg aatattctcg
     1321 ggcataaact cgagtacaac tacaattccc acaacgtgta catcactgcc gataagcaga
     1381 agaatgggat taaggcgaac ttcaagatcc gacacaacat cgaggatggt ggggtccagc
     1441 ttgccgacca ttatcagcag aacacaccca ttggggatgg accagtcctg ttgccggata
     1501 accactacct cagttaccag agcaaactgt ccaaagatcc caatgagaag agggaccata
     1561 tggtactgct cgaattcgtg actgcagcag gcattaccct tgggatggat gaactttata
     1621 agtcaggcta ataacagctt ccggactcta gaacatccct acaggtgata tcctcgggta
     1681 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1741 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1801 atcatgtctg tcgtcgaatt cgagttggcg cgcctgtcat tctaaatctc tctttcagcc
     1861 taaagctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt
     1921 tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg
     1981 gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc
     2041 ccctagcggg ggagggacgt aattacatcc ctgggggctt tgggaggggg ctgtccccgt
     2101 gagctcccag gcgcgcctgt cattctaaat ctctctttca gcctaaagct ttttccccgt
     2161 atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc
     2221 cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg
     2281 gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga
     2341 cgtaattaca tccctggggg ctttgggggg gggctgtccc cgtgagctcc caggcgcgcc
     2401 aactcggtac ctcacccgat tctcatcatg gagacacatc aggacatctc caataccgcc
     2461 atggaaacgc accaagatgc gactgctctt ctgccagtga cggcattcgt ctgcaagatt
     2521 aacggcatac agaaagaaac gatgagaccc attcaggacg agcctcagac tccagcgtaa
     2581 ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat
     2641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag
     2761 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2821 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2881 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2941 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3001 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3061 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3181 cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3241 aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct
     3301 cattaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3361 atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc
     3421 cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata
     3481 aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc
     3541 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3601 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3661 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3721 gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca
     3781 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt
     3841 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3901 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3961 ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga
     4021 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4081 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4141 acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca
     4201 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     4261 cgatcagtca aaagcctccg gtcggaggct tttga
//
