LOCUS       sequence-501949-        7917 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Express ULK1-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 7917)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7917)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: ULK1-NanoLuc Fusion Vector_
FEATURES             Location/Qualifiers
     source          1..7917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2324..2343)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2325..2343)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2385..2404)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2387..2519
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2655..2858)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2684..2704)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2859..3238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    3429..3712
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      3828..4416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4317..4336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4655..4703
                     /note="synthetic polyadenylation signal"
     CDS             complement(4767..5561)
                     /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(4878..4897)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5488..5507
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5593..5640)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5655..6012)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5669..5804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5821..6012
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5969..5989
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    6257..6378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6294..6313)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6348..6367
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6415..6433
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(6521..7033)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
ORIGIN
        1 gcgtgctccc gctcggggga gagcccacgg tgaagaccac cggggagggg ctgctggtgc
       61 ccccagcacg ggctcctggg tgcaggctcc ctccaaagcc agctgagggt gcgatctcca
      121 tgggcttctc ctgcaggctc tccgtgctgc ccgggtccgg ggcttgtgtc ccaaacgccg
      181 ccttaaggag caggtcagtg aggcggctgg tgctgaaaga ccggccaaag gggcccttgg
      241 ggtcctcgcc tggccgcagg ccagggccca acggctgacc atggaagggt cccacctccg
      301 agaggtcggg cagcgtccgg tttcgagggg gcgtcatcac cacgccctgc cgggctagga
      361 gggccagcag gttctgggag ctgggggtct tcgggaagtc aaaggagggc acagccttgg
      421 tgggggagcc caggatgggg ggcagggggt ttcgctgcag gaagtcgggc agcttgggtg
      481 agccccgcag gttccggcag gactgcaggc cgtgggacgg ctggggaggg ctggcctgtg
      541 gtgggctgaa cacagctccc agggggtccg tggggggttt gggcagcttg gggcggacga
      601 cgtgcaagtc agacaggttg ggggcgctgt gcaggcggca gcccagcccg gaagtgcggg
      661 gagagtgctc gggtgcagag gagcctggac gaggggacct gccaccccgc atctcagctc
      721 cctgtgggga gggcgtcccg ctccagcctg gccgctcagg gatggttcca acttgaggag
      781 atggcgtgta gggccggcct ccacccagag acatcttcct ggccaggacg cctccatgct
      841 cagcgtgggc agggggcgag gggctggccc ttgcaaagcc cagggggctg gtgctgcctg
      901 acctgcggat ggcagaggac cgaggtgttt ggaactgggt gggtgactgc aggtttcgct
      961 caatgcgctg gtagttctgc acctgcgtgg ggactgggat ggggacagag gcgccgcacc
     1021 tgctgctgga gaacgggcca gcccggcctg agggactggg ggagctgctg cagggtgggg
     1081 atggagatgg ggtccggccg tggctctcca agcccgcaga ggccaccagt gagctcccac
     1141 tgcacatcag gctgtctggc gggggtttgg cactgggcgc ctcagccacc aggtcacctg
     1201 gaaactgcgc ggggaccatg acgaagtcgt ctgtgtcaca ggaagagtcc ttgctgccac
     1261 cagagtcccg ggagctgtgc aggaagccag cggtgtcagc cggggaggcc agggtcttct
     1321 gcagctgctg catctcgccc agggacggcg gggaggccag gtgggaggtg gagctgctgc
     1381 tggagctgct gccggacccc gagcttgggt acgagggcac aggcacgggt ggggatttcc
     1441 tgaccgaggg gctggcatcg aggaaagggt gatgaaaaaa ctcatcgaag tccatgcggt
     1501 ccttgtggtt gcgttgcagt agggccagga gcagctgccg cagcggggcc gaggtctccc
     1561 gggggatggt ggggaccaac gtcttgttct tctcgtagaa caggcgcagg tcctgggggc
     1621 tgctggcctg gaagggcgcc ttccccgtca ggcactggta gacgatggtg ccgatgctcc
     1681 acaggtccgc cttcccgtcg tagtgctggg acatgatgac ctcgggggcc atgtacatgg
     1741 gggagccgca gagtgtggcc gccatcatgt tgctctggag gtaccgcgcg aagccgaagt
     1801 cagcgatctt gacgcggatg ctgttggggt tggcgcggcg gccggcgggg ttggacagca
     1861 ggatgttctg cggtttcagg tcgcggtgga tgatgccttt gctgtgcaga agccgcatgg
     1921 cgcccgcgat ctgctgcagg aagagcctga tggtgtcctc gctcagcgtg cgcatggcgt
     1981 gcaggtagtc ggccaggtcc ccaccgttgc agtactccat aaccaggtag acagaattag
     2041 ccatttcctg gaagtcgtac agggccacga tgttttcatg tttcagttcc ttcaggattt
     2101 tgatttcctt ccccagcagc gtctgagact tggcgaggtt cttcttgtta atgcacttga
     2161 cggcgacctc caaatcgtgc ttctcgcggt ggcggccctt gaagaccacc gcgaaggcgc
     2221 cgtggccgat caggtccttg cgggagaact cgaacttgcc cacggtctct gtgccgccgc
     2281 ggccgggctc catggttcta tctccttcga aagcgatcgc tagccctata gtgagtcgta
     2341 ttaatactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
     2401 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
     2461 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
     2521 tgcccagtgc ctcacgacca acttctgcag cttaagttcg agactgttgt gtcagaagca
     2581 ctgactgcgt tagcaattta actgtgataa actaccgcaa taaagcttct agtgatctga
     2641 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     2701 tgcgtcaacg gggcggggtt attacgacat tttggaaagt cccgttgatt ttggtgccaa
     2761 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     2821 tatccacgcc cattggtgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     2881 acgtagatgt actgccaagt aggaaagtcc cgtaaggtca tgtactgggc ataatgccag
     2941 gcgggccatt taccgtcatt gacgtcaata gggggcggac ttggcatatg atacacttga
     3001 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     3061 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     3121 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     3181 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgccaa
     3241 catggcggtc atattggaca tgagccaata taaatgtaca tattatgata tagatacaac
     3301 gtatgcaatg gccaatagcc aatattgatt tatgctatat aaccaatgaa taatatggct
     3361 aatggccaat attgaagatc tcgatcctct acgccggacg catcgtggcc ggagtactgc
     3421 ggtttatctc gatggctacg agggcagaca gtaagtggat ttaccataat cccttaattg
     3481 tacgcaccgc taaaacgcgt tcagcgcgat cacggcagca gacaggtaaa aatggcaaca
     3541 aaccacccga aaaactgccg caatcgcgcc tgataaattt taaccgtatg aatacctatg
     3601 caaccagagg gtacaggcca cattaccccc acttaatcca ctgaagctgc catttttcat
     3661 ggtttcacca tcccagcgaa gggccatcca gcgtgcgttc ctgtatttcc ggactataaa
     3721 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3781 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg aaatcctttt
     3841 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3901 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3961 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4021 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4081 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4141 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4201 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4261 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4321 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4381 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4441 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4501 tctgtggata accgtattac cgcctttgag tgagctgata ccgggtagaa agccagtccg
     4561 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     4621 gcaagcgcaa agaggatcct tatcgctatc gattcacaca aaaaaccaac acacagatgt
     4681 aatgaaaata aagatatttt attgcggcca tcgtgatggc aggttgggcg tcgcttggtc
     4741 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     4801 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4861 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     4921 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4981 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     5041 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     5101 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     5161 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     5221 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     5281 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     5341 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     5401 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     5461 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     5521 gcggccggag aacctgcgtg caatccatct tgttcaatca tgatctgcct cctgggttta
     5581 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     5641 agttaattaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5701 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     5761 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     5821 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     5881 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     5941 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     6001 tttccgcctc agaaggtacc taaccaagtt cctctttcag aggttatttc aggccatggt
     6061 gctgcgcaga tccgcgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6121 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gattttacca catttgtaga
     6181 ggttttactt gcttaaaaaa acctcccaca tctccccctg aacctgaaac ataaaatgaa
     6241 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6301 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6361 actcatcaat gtatcttatc atgtctgctc gaagcggccg ccccaagggg ttatgctagt
     6421 tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga
     6481 tcagcttgca tgcctgcagg tcgactctag agtcgcggcc ttacgccaga atgcgttcgc
     6541 acagccgcca gccggtcact ccgttgatgg ttactcggaa cagcagggag ccgtcggggt
     6601 tgatcaggcg ctcgtcgata attttgttgc cgttccacag ggtccctgtt acagtgatct
     6661 ttttgccgtc gaacacggcg atgccttcat acggccgtcc gaaatagtcg atcatgttcg
     6721 gcgtaacccc gtcgattacc agtgtgccat agtgcaggat caccttaaag tgatgatcat
     6781 ccacagggta caccacctta aaaatttttt cgatctggcc catttggtcg ccgctcagac
     6841 cttcatacgg gatgatgaca tggatgtcga tcttcagccc attttcaccg ctcaggacaa
     6901 tcctttggat cggagttacg gacaccccga gattctgaaa caaactggac acacctccct
     6961 gttcaaggac ttggtccagg ttgtagccgg ctgtctgtcg ccagtcccca acgaaatctt
     7021 cgagtgtgaa gacgccgctc gagccgagag aaactacgta ggcacagatg ccagtcagca
     7081 gcgccgagag tctccgctca atgcacagct tgcacttggt gacgttctcg atgtcggcct
     7141 ggtccgagag catgtgctgc agcccctcca ggagcagcag ggccttgtgg tagcgtggga
     7201 cgcagccctc acggtgctgg aacatctcgt ccagggcagc cgactgcacc atctgcacag
     7261 cgtggctgaa gatgagcctc tcggcagtga tgctgtgaat gcggtccagg agccgctgct
     7321 tgtccaggaa gaagcgctgc agccgcaggc tcaggccctg gcaggacacc acgctggcct
     7381 tgtacagctc attcagcctg cgcaccacct gcttcacagt ggacgacagg cagagcttgc
     7441 cggcccggat ctggtcgatg gcactttgca ggccggagga cagtagctcg gccaccttca
     7501 ggtacagcac cagctgttcc gcgaagcccc attctcggct cagcaggctg atctggtcgg
     7561 ccaccacact ctcctgcagc tggtactcag ggccccccgc cgcctcactg gcgctgccct
     7621 tcagggctgc gatctccagg acgtgctgca cgaacagcag cgtgaagcgc aggccacgca
     7681 ggatctccgt gtgctcttgc tccatgaggg tctcctcagg gaggtcgggg gcctcgaagg
     7741 tcacagcccc ctccaggttc gcagcaatgg ggtcggcaaa gctgcagccg tgtcctggag
     7801 cagggagctc aggggctggg gcctcgctgc agggcccagg caccaggtgg cgggcagaag
     7861 agctggcaga gccagtgggg cccgctgaga acatcctggt gcgggggccc tgggggg
//
