LOCUS       sequence-480605-        5682 bp    DNA     circular SYN 25-JUL-2025
DEFINITION  Express PIM1-NanoLuc Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5682)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: PIM1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2025..2537
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2625..2643)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2680..2801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2691..2710)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2745..2764
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3046..3403
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3046..3237
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3069..3089)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3254..3389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3418..3465
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3497..4291
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3551..3570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4161..4180
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4355..4403
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4642..5230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4722..4741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5346..5629
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgctc ttgtccaaaa
     1081 tcaactcgct tgcccacctg cgcgccgcgc cctgcaacga cctgcacgcc accaagctgg
     1141 cgcccggcaa ggagaaggag cccctggagt cgcagtacca ggtgggcccg ctactgggca
     1201 gcggcggctt cggctcggtc tactcaggca tccgcgtctc cgacaacttg ccggtggcca
     1261 tcaaacacgt ggagaaggac cggatttccg actggggaga gctgcctaat ggcactcgag
     1321 tgcccatgga agtggtcctg ctgaagaagg tgagctcggg tttctccggc gtcattaggc
     1381 tcctggactg gttcgagagg cccgacagtt tcgtcctgat cctggagagg cccgagccgg
     1441 tgcaagatct cttcgacttc atcacggaaa ggggagccct gcaagaggag ctggcccgca
     1501 gcttcttctg gcaggtgctg gaggccgtgc ggcactgcca caactgcggg gtgctccacc
     1561 gcgacatcaa ggacgaaaac atccttatcg acctcaatcg cggcgagctc aagctcatcg
     1621 acttcgggtc gggggcgctg ctcaaggaca ccgtctacac ggacttcgat gggacccgag
     1681 tgtatagccc tccagagtgg atccgctacc atcgctacca tggcaggtcg gcggcagtct
     1741 ggtccctggg gatcctgctg tatgatatgg tgtgtggaga tattcctttc gagcatgacg
     1801 aagagatcat caggggccag gttttcttca ggcagagggt ctcttcagaa tgtcagcatc
     1861 tcattagatg gtgcttggcc ctgagaccat cagataggcc aaccttcgaa gaaatccaga
     1921 accatccatg gatgcaagat gttctcctgc cccaggaaac tgctgagatc cacctccaca
     1981 gcctgtcgcc ggggcccagc aaagtttctc tcggctcgag cggcgtcttc acactcgaag
     2041 atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg
     2101 gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg
     2161 tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc
     2221 tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg
     2281 atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga
     2341 acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga
     2401 tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc
     2461 ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg
     2521 aacgcattct ggcgtaaggc cgcgactcta gagtcgacct gcaggcatgc aagctgatcc
     2581 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2641 agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2701 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2761 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2821 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct
     2881 acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     2941 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     3001 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     3061 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3121 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3241 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3301 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3361 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt
     3421 gacaattaat catcggcata gtatgtcggc atagtataat acgacaaggt gaggaactaa
     3481 acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3541 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3601 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3661 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3721 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3781 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3841 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3901 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3961 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4021 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4081 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4141 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4201 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4261 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4321 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     4381 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt
     4441 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     4501 actggctttc tacccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5041 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5221 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5341 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     5401 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     5461 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     5521 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     5581 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     5641 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
