LOCUS       sequence-249560-        4361 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 4361)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4361)
  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-001   LifeAct-Sirius//BsaI
FEATURES             Location/Qualifiers
     source          1..4361
                     /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    1746..1867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1783..1802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1837..1856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      2668..2697
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2719..3306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3207..3226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3318..3345)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3369..4160,4161..4229))
                     /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     3992..4011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4230..4332)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4332..4361,1..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 gagcaaagga gaggagctct ttacaggagt agtcccgata ttggtggaac
      961 ttgatggcga cgtcaatgga caccgtttct ccgtgagtgg agaaggcgaa ggcgatgcaa
     1021 cctatggtaa actcaccctg aagcttatct gcactacggg gaaactgcca gtaccttggc
     1081 caaccctggt tacaaccctg cagtttgggg ttctgtgttt tgcccgatat cccgatcaca
     1141 tgaagcagca tgacttcttc aagtcagcca tgccagaagg gtatgtccaa gagaggacca
     1201 tcttctttaa agacgatggc aactataaga cgagagcaga agtgaagttt gagggagaca
     1261 ctctggtgaa tcggattgaa ctcaaaggca tcgacttcaa ggaagatggc aatatcctgg
     1321 gtcacaaact ggagtacaat gggatctcta gcaacgtgta catcacagcc gataagcaga
     1381 agaacgggat aaaggcgcat ttcaagattc gccataacat cgaggacggt ggcgttcaac
     1441 tcgctgatca ctaccagcag aacactccca ttggcgatgg acctgtgctt ctccctgaca
     1501 accactactt gtctgtccag agcaagctgt ccaaagaccc caatgagaag agggaccata
     1561 tggtcttgct ggagagtgtg acagctgctg ggattactct gggtatggac gagctgtaca
     1621 aatcaggctc tgattaaggc gccaggtaca tctccaatac ggcgtcttgc atagagaaca
     1681 cgattagcac accggttacc cacgtgatgc ccgaacggac ccatgtcatg ccagagcgca
     1741 ctggtaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1801 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1861 tatcttatca tgtctgtcgt cgaattcgag ttggcgcgcc tgtcattcta aatctctctt
     1921 tcagcctaaa gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     1981 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     2041 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     2101 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg agggggctgt
     2161 ccccgtgagc tcccaggcgc gcctgtcatt ctaaatctct ctttcagcct aaagcttttt
     2221 ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag
     2281 cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg
     2341 cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg
     2401 gagggacgta attacatccc tgggggcttt gggggggggc tgtccccgtg agctcccagg
     2461 cgcgccaact cggtacctca cccgattctc atcatggaga cacatcagga catctccaat
     2521 accgccatgg aaacgcacca agatgcgact gctcttctgc cagtgacggc attcgtctgc
     2581 aagattaacg gcatacagaa agaaacgatg agacccattc aggacgagcc tcagactcca
     2641 gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct ttcttcggta
     2701 gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2761 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2821 tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2881 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2941 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3001 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3061 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3121 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3181 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3241 tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta
     3301 tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacatttg
     3361 ggccctcatt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3421 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca
     3481 tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca
     3541 gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc
     3601 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3661 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3721 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3781 aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg
     3841 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga
     3901 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3961 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4021 aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg
     4081 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4141 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4201 agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat
     4261 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     4321 acgtgccgat cagtcaaaag cctccggtcg gaggcttttg a
//
