LOCUS       sequence-473363-        5712 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express PP1Cg-NanoLuc(R) 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 5712)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5712)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PP1Cg-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5712
                     /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             2055..2567
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2655..2673)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2710..2831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2721..2740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2775..2794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3076..3433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3076..3267
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3099..3119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3284..3419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3448..3495
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3527..4321
                     /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(3581..3600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4191..4210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4385..4433
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4672..5260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4752..4771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5376..5659
                     /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 cgccatggcg gatttagata
     1081 aactcaacat cgacagcatt atccaacggc tgctggaagt gagagggtcc aagcctggta
     1141 agaatgtcca gcttcaggag aatgaaatca gaggactgtg cttaaagtct cgtgaaatct
     1201 ttctcagtca gcctatccta ctagaacttg aagcaccact caaaatatgt ggtgacatcc
     1261 atggacaata ctatgatttg ctgcgacttt ttgagtacgg tggtttccca ccagaaagca
     1321 actacctgtt tcttggggac tatgtggaca ggggaaagca gtcattggag acgatctgcc
     1381 tcttactggc ctacaaaata aaatatcctg agaatttttt tcttctcaga gggaaccatg
     1441 aatgtgccag catcaacaga atttatggat tttatgatga atgtaaaaga agatacaaca
     1501 ttaaactatg gaaaactttc acagactgtt ttaactgttt accgatagca gccatcgtgg
     1561 atgagaagat attctgctgt catggaggtt tatcaccaga tcttcaatct atggagcaga
     1621 ttcggcgaat tatgcgacca actgatgtac cagatcaagg tcttctttgt gatcttttgt
     1681 ggtctgaccc cgataaagat gtcttaggct ggggtgaaaa tgacagagga gtgtccttca
     1741 catttggtgc agaagtggtt gcaaaatttc tccataagca tgatttggat cttatatgta
     1801 gagcccatca ggtggttgaa gatggatatg aattttttgc aaagaggcag ttggtcactc
     1861 tgttttctgc gcccaattat tgcggagagt ttgacaatgc aggtgccatg atgagtgtgg
     1921 atgaaacact aatgtgttct tttcagattt taaagcctgc agagaaaaag aagccaaatg
     1981 ccacgagacc tgtaacgcct ccaaggggta tgatcacaaa gcaagcaaag aaagtttctc
     2041 tcggctcgag cggcgtcttc acactcgaag atttcgttgg ggactggcga cagacagccg
     2101 gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg
     2161 gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg
     2221 acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa
     2281 aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg
     2341 gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg
     2401 aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca
     2461 acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca
     2521 tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcgtaaggc cgcgactcta
     2581 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     2641 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag
     2701 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2761 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2821 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     2881 gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa
     2941 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3001 cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac
     3061 ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3181 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3241 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3301 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3361 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3421 aggcttttgc aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc
     3481 atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga
     3541 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3601 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3661 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3721 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3781 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3841 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3901 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3961 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4021 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4081 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4141 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4201 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4261 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4321 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     4381 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4441 agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     4501 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacccggtat cagctcactc
     4561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5101 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5221 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     5281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5341 atcaaaaagg atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg
     5401 atggcccttc gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg
     5461 ggtaatgtgg cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc
     5521 gcgattgcgg cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc
     5581 gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt
     5641 ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga
     5701 ggatcgagat ct
//
