LOCUS       sequence-436830-        3211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PAB1-5'UTR-NLuc mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3211)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 3211)
  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: pUC57-T7-PAB1-NLuc
FEATURES             Location/Qualifiers
     source          1..3211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     402..421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        402..420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1121..1137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1121..1137)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1134..1156)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1145..1161
                     /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(1169..1175,1176..1193,1194..1199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1214..1235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1352..1369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1523..2111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1603..1622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2289..3098)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3012..3031
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3090..3109
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt ctaatacgac tcactatagg
      421 gagtaacatt tttacttaat aaccgtttga agcatagagc aggccctggt accaccacct
      481 aatatctggc tttttattca ataaaaactc aaaaaaaaaa atccaaaaaa aactaaaaaa
      541 ccaataaaaa taaaatggtt tttactttag aagattttgt tggtgattgg cgtcaaactg
      601 caggttataa tttagatcaa gttttagaac aaggtggtgt ttcttctttg tttcaaaact
      661 taggtgtttc tgttactcct attcaacgta ttgttttatc tggtgaaaat ggtttgaaga
      721 ttgatattca tgttattatt ccatatgaag gtttgtctgg tgatcaaatg ggtcaaattg
      781 aaaagatttt caaagttgtt tatcctgtcg acgatcatca tttcaaagtt attttgcatt
      841 atggtacttt agttattgat ggtgttactc ctaatatgat tgattatttt ggtcgtcctt
      901 atgaaggtat tgctgttttc gatggtaaaa agatcactgt tactggtact ttatggaatg
      961 gtaacaagat tattgatgaa cgtttgatta atcctgatgg ttctttatta tttcgtgtta
     1021 ctattaatgg tgttactggt tggcgtttat gtgaacgtat tttagcttaa ctcgagtcat
     1081 gtaattagtt ggatccgcat gcaagcttgg tgtaatcatg gtcatagctg tttcctgtgt
     1141 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1201 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1261 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1321 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1381 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1441 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1501 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1561 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1621 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1681 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1741 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1801 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1861 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1921 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1981 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2041 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2101 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2161 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2221 taaattaaaa atgaagtttt aaatcaagcc caatctgaat aatgttacaa ccaattaacc
     2281 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     2341 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2401 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2461 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     2521 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     2581 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     2641 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     2701 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     2761 tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac
     2821 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     2881 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     2941 ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat
     3001 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     3061 aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3121 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     3181 cagagatttt gagacacggg ccagagctgc a
//
