LOCUS       sequence-436835-        3178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  betaglobin-5'UTR-NanoLuc mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3178)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 3178)
  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_beta-globin-5'UTR_Nanoluc
FEATURES             Location/Qualifiers
     source          1..3178
                     /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"
     regulatory      490..499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1088..1104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1088..1104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1101..1123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1112..1128
                     /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(1136..1142,1143..1160,1161..1166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1181..1202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1319..1336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1490..2078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1570..1589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2256..3065)
                     /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     2979..2998
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3057..3076
                     /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 gagagagctc acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc
      481 ctgcaggccg ccaccatggt ttttacttta gaagattttg ttggtgattg gcgtcaaact
      541 gctggttata atttagatca agttttagaa caaggtggtg tttcttcttt gtttcaaaac
      601 ttaggtgttt ctgttactcc tattcaacgt attgttttat ctggtgaaaa tggtttgaag
      661 attgatattc atgttattat tccatatgaa ggtttgtctg gtgatcaaat gggtcaaatt
      721 gaaaagattt tcaaagttgt ttatcctgtg gacgatcatc atttcaaagt tattttgcat
      781 tatggtactt tagttattga tggtgttact cctaatatga ttgattattt tggtcgtcct
      841 tatgaaggta ttgctgtttt cgatggtaaa aagatcactg ttactggtac tttatggaat
      901 ggtaacaaga ttattgatga acgtttgatt aatcctgatg gttctttatt atttcgtgtt
      961 actattaatg gtgttactgg ttggcgttta tgtgaacgta ttttagctta agcggccgca
     1021 ccggtgctag ctcgagacta gtcgacaaaa aaaaaaaaaa aaaaaaaaaa agcttggtgt
     1081 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1141 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1201 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1921 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaagcccaa
     2221 tctgaataat gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     2281 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2341 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2401 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2461 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2521 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2581 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2641 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2701 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2761 tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2821 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2881 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2941 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3001 tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg
     3061 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     3121 atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgca
//
