LOCUS       sequence-273530-        4257 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 4257)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 4257)
  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-032   Lamp1-Eos//BsaI
FEATURES             Location/Qualifiers
     source          1..4257
                     /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    1641..1762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1678..1697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1732..1751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      2563..2592
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2614..3201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3102..3121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3213..3240)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(3264..4055,4056..4124))
                     /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     3887..3906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4125..4227)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4228..4257,1..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 ggggcgrggg 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 aataacagct tccggactct agaacatccc
     1621 tacaggtgat atcctcgggt aacttgttta ttgcagctta taatggttac aaataaagca
     1681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1741 ccaaactcat caatgtatct tatcatgtct gtcgtcgaat tcgagttggc gcgcctgtca
     1801 ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc
     1861 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     1921 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     1981 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     2041 ttgggagggg gctgtccccg tgagctccca ggcgcgcctg tcattctaaa tctctctttc
     2101 agcctaaagc tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa
     2161 gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct
     2221 gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc
     2281 tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc
     2341 ccgtgagctc ccaggcgcgc caactcggta cctcacccga ttctcatcat ggagacacat
     2401 caggacatct ccaataccgc catggaaacg caccaagatg cgactgctct tctgccagtg
     2461 acggcattcg tctgcaagat taacggcata cagaaagaaa cgatgagacc cattcaggac
     2521 gagcctcaga ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg
     2581 ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2701 gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2761 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2821 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2881 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2941 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3001 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3061 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3121 tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca
     3181 ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc
     3241 aggtgattac atttgggccc tcattaccaa tgcttaatca gtgaggcacc tatctcagcg
     3301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg
     3361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg
     3421 gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct
     3481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt
     3721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3781 atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca
     3961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct
     4141 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     4201 aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttga
//
