LOCUS       sequence-298160-        4377 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 4377)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4377)
  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..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      196..205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             202..252
                     /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    1123..1244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1160..1179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1214..1233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      2045..2074
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2096..2683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2584..2603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2695..2722)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2746..3537,3538..3606))
                     /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     3369..3388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3607..3709)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3710..3741)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     enhancer        3897..4276
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4278..4377,1..176)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggcgggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
       61 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      121 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      181 tcgctgcgtt gggacgccac catgggcgtg gccgacctga tcaagaagtt cgagagcatc
      241 agcaaggagg agggcgggtc aggcggcagc gtcagcaaag gcgaggagct catcaaggag
      301 aacatgcaca ccaagctgta catggaaggt actgtgaata accaccactt caaatgcacg
      361 catgaaggtg aaggcaagcc ctatgagggc acacagacca atcgcatcaa agtggttgag
      421 ggaggtcctc tgccgtttgc gttcgacatt ctggccacat gcttcatgta tggctccaaa
      481 acgttcatca accacactca gggcataccc gatttcttca agcagtcttt tcctgaaggc
      541 tttacatggg agagagtgac cacatacgaa gatggaggcg tcctcactgt gacacaagac
      601 acaagtctgc aggatggatg tctcatctac aatgtgaagc tgcgtggggt gaactttccc
      661 tctaatgggc ctgtaatgca gaagaaaact cttgggtggg aagcatcaac cgagactctt
      721 tacccagctg atggagggct tgaaggccgg tgtgacatgg ctctcaagtt ggtaggtgga
      781 ggacatctgc attgcaacct gaaaaccacc tacaggagca agaagccagc aaagaacctg
      841 aagatgccag gagtctattt tgtggacaga cgattggagc ggattaagga ggccgacaat
      901 gaaacctatg tcgagcaaca cgaggttgcc gttgctaggt attgtgactt gccttccaaa
      961 ctggggcata agctgaacgg gatggatgag ctgtacaaat caggctctga ttaaggcgcc
     1021 aggtacatct ccaatacggc gtcttgcata gagaacacga ttagcacacc ggttacccac
     1081 gtgatgcccg aacggaccca tgtcatgcca gagcgcactg gtaacttgtt tattgcagct
     1141 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     1201 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtcgtcga
     1261 attcgagttg gcgcgcctgt cattctaaat ctctctttca gcctaaagct ttttccccgt
     1321 atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc
     1381 cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg
     1441 gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga
     1501 cgtaattaca tccctggggg ctttgggagg gggctgtccc cgtgagctcc caggcgcgcc
     1561 tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc caggtgtctg
     1621 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt
     1681 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc
     1741 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg
     1801 gggctttggg ggggggctgt ccccgtgagc tcccaggcgc gccaactcgg tacctcaccc
     1861 gattctcatc atggagacac atcaggacat ctccaatacc gccatggaaa cgcaccaaga
     1921 tgcgactgct cttctgccag tgacggcatt cgtctgcaag attaacggca tacagaaaga
     1981 aacgatgaga cccattcagg acgagcctca gactccagcg taactggact gcaatcaact
     2041 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag
     2101 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2161 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag
     2221 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2281 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2341 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2401 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2461 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2521 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2581 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca
     2641 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga
     2701 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcattacc aatgcttaat
     2761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2821 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     2881 accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     2941 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3181 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3241 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     3301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3421 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3601 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     3661 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     3721 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa aggtctcata
     3781 ggtcttgtat tcagtctcgc actttagaca tggtcttctg ctgcccgtgg tcctctattc
     3841 gtcggatgtt ctgtcgattc ttcctgtcta tctggactgg atctgatatc atcgtcgaca
     3901 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     3961 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4021 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4081 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4141 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4201 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4261 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     4321 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgat
//
