LOCUS       sequence-475936-        2976 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  BsaI Golden Gate assembly plasmid encoding the EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2976)
  AUTHORS   Peterman EL, Ploessl DS, Love KS, Sanabria V, Daniels RF, Johnstone 
            CP, Godavarti DR, Kabaria SR, Oakes CG, Pai AA, Galloway KE
  TITLE     High-resolution profiling reveals coupled transcriptional and 
            translational regulation of transgenes.
  JOURNAL   Nucleic Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528
  PUBMED    39651241
REFERENCE   2  (bases 1 to 2976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528"
COMMENT     Sequence Label: pKG2019_pPV1-EF1a
FEATURES             Location/Qualifiers
     source          1..2976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..1230
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          279..1221
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1178..1198
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        1276..1380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1381..1449,1450..2241)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGYRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1599..1618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2412..2976,1..24)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2901..2920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaggggggc ggagcctatg gaaaaacgcc agcaacgggt ctcatactgc tccggtgccc
       61 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      121 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      181 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      241 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
      301 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacgcccc
      361 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      421 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcttgggcgc
      481 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      541 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      601 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      661 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      721 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
      781 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
      841 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
      901 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
      961 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1021 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1081 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1141 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1201 agtttttttc ttccatttca ggtgtcgtga gaatgagaga cctagacgtc aggtggcact
     1261 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1321 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1381 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1441 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1501 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1561 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1621 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1681 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1741 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1801 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1861 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1921 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1981 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2041 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtat
     2101 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2161 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2221 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2281 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2341 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2401 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2461 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2521 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2581 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2641 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2701 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2761 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2821 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2881 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2941 cacctctgac ttgagcgtcg atttttgtga tgctcg
//
