LOCUS       sequence-420629-        4813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4813)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4813)
  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: pCS2(plus)EGFP_H
FEATURES             Location/Qualifiers
     source          1..4813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..985
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1020..1038
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1020..1037
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1071..1080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1077..1079,1080..1082,1083..1793)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1077..1079,1080..1082,1083..1793)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1122..1143)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1383..1402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1730..1751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1850..1984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1874..1893)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1928..1947
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2097..2117)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2097..2115)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2136..2152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2136..2152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2149..2171)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2160..2176
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2184..2190,2191..2208,2209..2214))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2229..2250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2367..2384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2538..3126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2618..2637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3297..4088,4089..4157))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3920..3939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4158..4262)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4288..4743)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4425..4446)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4637..4656
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
       61 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
      121 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
      181 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
      241 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
      301 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
      361 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
      421 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
      481 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
      541 tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat
      601 tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg
      661 acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca
      721 atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt
      781 acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg
      841 gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat
      901 gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta
      961 tataagcaat gctcgtttag ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga
     1021 tttaggtgac actatagaat acaagctact tgttcttttt gcaggatccc gccaccatgg
     1081 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1141 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1201 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1261 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1321 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1381 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1441 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1501 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1561 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1621 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1681 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1741 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggag
     1801 aattcaaggc ctctcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     1861 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1921 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1981 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2041 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     2101 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2161 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     2221 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2281 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2341 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2401 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2461 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2521 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2581 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2641 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2701 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2761 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2821 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2881 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2941 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3001 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3061 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3121 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3181 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3241 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3301 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3361 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3421 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3481 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3541 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3601 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3661 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3721 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3781 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3841 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3901 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3961 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4021 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4081 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4141 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4201 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4261 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4321 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4381 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4441 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4501 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4561 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4621 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4681 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4741 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4801 cgctattacg cca
//
