LOCUS       sequence-273335-        4323 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 4323)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4323)
  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-008   LifeAct-Eos//BsaI
FEATURES             Location/Qualifiers
     source          1..4323
                     /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"
     polyA_signal    1707..1828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1744..1763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1798..1817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      2629..2658
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2680..3267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3168..3187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3279..3306)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3330..4121,4122..4190))
                     /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     3953..3972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4191..4293)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4294..4323,1..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 ggggcggggs ggggsgrggg 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 gcggcagctc agcgatcaaa cccgacatga agatcaacct ccgaatggaa ggcaacgtaa
      961 acggccatca cttcgtgatt gatggcgatg gcacagggaa gccatttgag gggaaacaaa
     1021 gcatggatct ggaggtgaag gaaggtggtc ctctgccttt tgccttcgac atactgacta
     1081 cagcctttca ctatgggaat cgcgtgtttg tcgagtatcc cgaccatata caggattact
     1141 tcaagcagag tttcccgaaa ggctatagct gggaaaggtc cctcacattt gaggatggag
     1201 ggatttgcat tgcaagaaac gacatcacca tggaaggaga caccttctac aacaaggttc
     1261 ggttccatgg gacaaatttc cccgctaatg gtcctgtgat gcagaagaaa accctgaagt
     1321 gggaaccatc cactgagaag atgtacgtga gagatggagt actgacgggt gacattcacg
     1381 ctgcactgct gttggagggc aatgcccact atcggtgtga ttttcgcact acctacaagg
     1441 ccaaagagaa aggcgtcaaa ctccctggat accaccttgt cgatcattgc atcgagatcc
     1501 tttctcacga caaggactac aacaaggtga agctgtatga acatgccgtt gctcactctg
     1561 gattgccaga caatgcaagg cgttcaggct ctgattaagg cgccaggtac atctccaata
     1621 cggcgtcttg catagagaac acgattagca caccggttac ccacgtgatg cccgaacgga
     1681 cccatgtcat gccagagcgc actggtaact tgtttattgc agcttataat ggttacaaat
     1741 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1801 gtttgtccaa actcatcaat gtatcttatc atgtctgtcg tcgaattcga gttggcgcgc
     1861 ctgtcattct aaatctctct ttcagcctaa agctttttcc ccgtatcccc ccaggtgtct
     1921 gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg
     1981 tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag
     2041 cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg
     2101 ggggctttgg gagggggctg tccccgtgag ctcccaggcg cgcctgtcat tctaaatctc
     2161 tctttcagcc taaagctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag
     2221 cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg
     2281 cacgctgccg gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct
     2341 cgctgctgcc ccctagcggg ggagggacgt aattacatcc ctgggggctt tggggggggg
     2401 ctgtccccgt gagctcccag gcgcgccaac tcggtacctc acccgattct catcatggag
     2461 acacatcagg acatctccaa taccgccatg gaaacgcacc aagatgcgac tgctcttctg
     2521 ccagtgacgg cattcgtctg caagattaac ggcatacaga aagaaacgat gagacccatt
     2581 caggacgagc ctcagactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca
     2641 cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg
     2701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2761 tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat
     2821 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2881 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2941 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3001 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3061 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc
     3241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg
     3301 tgagccaggt gattacattt gggccctcat taccaatgct taatcagtga ggcacctatc
     3361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3421 acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc
     3481 tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt
     3541 gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc
     3781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3841 actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc
     3901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa
     4021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc
     4201 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     4261 tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt
     4321 tga
//
