LOCUS       sequence-481502-        6581 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express NanoLuc-cIAP2 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 6581)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6581)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-cIAP2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6581
                     /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             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3524..3542)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3579..3700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3590..3609)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3644..3663
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3945..4302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3945..4136
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3968..3988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4153..4288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4317..4364
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4396..5190
                     /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(4450..4469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5060..5079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5254..5302
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5541..6129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5621..5640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6245..6528
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat gaacatagta gaaaacagca
     1621 tattcttatc aaatttgatg aaaagcgcca acacgtttga actgaaatac gacttgtcat
     1681 gtgaactgta ccgaatgtct acgtattcca cttttcctgc tggggttcct gtctcagaaa
     1741 ggagtcttgc tcgtgctggt ttctattaca ctggtgtgaa tgacaaggtc aaatgcttct
     1801 gttgtggcct gatgctggat aactggaaaa gaggagacag tcctactgaa aagcataaaa
     1861 agttgtatcc tagctgcaga ttcgttcaga gtctaaattc cgttaacaac ttggaagcta
     1921 cctctcagcc tacttttcct tcttcagtaa caaattccac acactcatta cttccgggta
     1981 cagaaaacag tggatatttc cgtggctctt attcaaactc tccatcaaat cctgtaaact
     2041 ccagagcaaa tcaagatttt tctgccttga tgagaagttc ctaccactgt gcaatgaata
     2101 acgaaaatgc cagattactt acttttcaga catggccatt gacttttctg tcgccaacag
     2161 atctggcaaa agcaggcttt tactacatag gacctggaga cagagtggct tgctttgcct
     2221 gtggtggaaa attgagcaat tgggaaccga aggataatgc tatgtcagaa cacctgagac
     2281 attttcccaa atgcccattt atagaaaatc agcttcaaga cacttcaaga tacacagttt
     2341 ctaatctgag catgcagaca catgcagccc gctttaaaac attctttaac tggccctcta
     2401 gtgttctagt taatcctgag cagcttgcaa gtgcgggttt ttattatgtg ggtaacagtg
     2461 atgatgtcaa atgcttttgc tgtgatggtg gactcaggtg ttgggaatct ggagatgatc
     2521 catgggttca acatgccaag tggtttccaa ggtgtgagta cttgataaga attaaaggac
     2581 aggagttcat ccgtcaagtt caagccagtt accctcatct acttgaacag ctgctatcca
     2641 catcagacag cccaggagat gaaaatgcag agtcatcaat tatccatttt gaacctggag
     2701 aagaccattc agaagatgca atcatgatga atactcctgt gattaatgct gccgtggaaa
     2761 tgggctttag tagaagcctg gtaaaacaga cagttcagag aaaaatccta gcaactggag
     2821 agaattatag actagtcaat gatcttgtgt tagacttact caatgcagaa gatgaaataa
     2881 gggaagagga gagagaaaga gcaactgagg aaaaagaatc aaatgattta ttattaatcc
     2941 ggaagaatag aatggcactt tttcaacatt tgacttgtgt aattccaatc ctggatagtc
     3001 tactaactgc cggaattatt aatgaacaag aacatgatgt tattaaacag aagacacaga
     3061 cgtctttaca agcaagagaa ctgattgata cgattttagt aaaaggaaat attgcagcca
     3121 ctgtattcag aaactctctg caagaagctg aagctgtgtt atatgagcat ttatttgtgc
     3181 aacaggacat aaaatatatt cccacagaag atgtttcaga tctaccagtg gaagaacaat
     3241 tgcggagact acaagaagaa agaacatgta aagtgtgtat ggacaaagaa gtgtccatag
     3301 tgtttattcc ttgtggtcat ctagtagtat gcaaagattg tgctccttct ttaagaaagt
     3361 gtcctatttg taggagtaca atcaagggta cagttcgtac atttctttca gtttaaacga
     3421 attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg
     3481 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3541 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3601 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3661 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3721 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3781 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     3841 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     3901 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3961 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4021 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4261 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg
     4321 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4381 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4441 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4501 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4561 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4621 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4681 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4741 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4801 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4861 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4921 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4981 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5041 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5101 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5161 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5221 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     5281 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt
     5341 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     5401 ctggctttct acccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5941 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6121 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6241 tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga
     6301 aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata
     6361 ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt
     6421 gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat
     6481 taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag
     6541 tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
