LOCUS       sequence-249569-        4376 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 4376)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4376)
  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-010   LifeAct-Neptune//BsaI
FEATURES             Location/Qualifiers
     source          1..4376
                     /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    1761..1882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1798..1817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1852..1871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      2683..2712
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2734..3321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3222..3241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3333..3360)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3384..4175,4176..4244))
                     /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     4007..4026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4245..4347)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4347..4376,1..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 cagcaaaggc gaggagctca tcaaggagaa catgcacacc aagctgtaca
      961 tggaaggtac tgtgaataac caccacttca aatgcacgca tgaaggtgaa ggcaagccct
     1021 atgagggcac acagaccaat cgcatcaaag tggttgaggg aggtcctctg ccgtttgcgt
     1081 tcgacattct ggccacatgc ttcatgtatg gctccaaaac gttcatcaac cacactcagg
     1141 gcatacccga tttcttcaag cagtcttttc ctgaaggctt tacatgggag agagtgacca
     1201 catacgaaga tggaggcgtc ctcactgtga cacaagacac aagtctgcag gatggatgtc
     1261 tcatctacaa tgtgaagctg cgtggggtga actttccctc taatgggcct gtaatgcaga
     1321 agaaaactct tgggtgggaa gcatcaaccg agactcttta cccagctgat ggagggcttg
     1381 aaggccggtg tgacatggct ctcaagttgg taggtggagg acatctgcat tgcaacctga
     1441 aaaccaccta caggagcaag aagccagcaa agaacctgaa gatgccagga gtctattttg
     1501 tggacagacg attggagcgg attaaggagg ccgacaatga aacctatgtc gagcaacacg
     1561 aggttgccgt tgctaggtat tgtgacttgc cttccaaact ggggcataag ctgaacggga
     1621 tggatgagct gtacaaatca ggctctgatt aaggcgccag gtacatctcc aatacggcgt
     1681 cttgcataga gaacacgatt agcacaccgg ttacccacgt gatgcccgaa cggacccatg
     1741 tcatgccaga gcgcactggt aacttgttta ttgcagctta taatggttac aaataaagca
     1801 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1861 ccaaactcat caatgtatct tatcatgtct gtcgtcgaat tcgagttggc gcgcctgtca
     1921 ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc
     1981 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     2041 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     2101 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     2161 ttgggagggg gctgtccccg tgagctccca ggcgcgcctg tcattctaaa tctctctttc
     2221 agcctaaagc tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa
     2281 gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct
     2341 gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc
     2401 tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc
     2461 ccgtgagctc ccaggcgcgc caactcggta cctcacccga ttctcatcat ggagacacat
     2521 caggacatct ccaataccgc catggaaacg caccaagatg cgactgctct tctgccagtg
     2581 acggcattcg tctgcaagat taacggcata cagaaagaaa cgatgagacc cattcaggac
     2641 gagcctcaga ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg
     2701 ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2821 gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3241 tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca
     3301 ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc
     3361 aggtgattac atttgggccc tcattaccaa tgcttaatca gtgaggcacc tatctcagcg
     3421 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg
     3481 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg
     3541 gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct
     3601 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3661 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3721 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3781 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt
     3841 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3901 atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3961 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4021 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca
     4081 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4141 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4201 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct
     4261 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     4321 aagttggaac ctcttacgtg ccgatcagtc aaaagcctcc ggtcggaggc ttttga
//
