LOCUS       sequence-471872-        6487 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express NanoLuc(R)-cIAP1 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 6487)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-cIAP1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6487
                     /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(3566..3584)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3621..3742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3632..3651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3686..3705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3987..4344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3987..4178
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4010..4030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4195..4330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4359..4406
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4438..5232
                     /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(4492..4511)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5102..5121
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5296..5344
                     /note="synthetic polyadenylation signal"
     rep_origin      5501..6046
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5947..5966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6163..6180
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 gcacaaaact gcctcccaaa
     1621 gacttttccc aggtccctcg tatcaaaaca ttaagagtat aatggaagat agcacgatct
     1681 tgtcagattg gacaaacagc aacaaacaaa aaatgaagta tgacttttcc tgtgaactct
     1741 acagaatgtc tacatattca actttccccg ccggggtgcc tgtctcagaa aggagtcttg
     1801 ctcgtgctgg tttctattat actggtgtga atgacaaggt caaatgcttc tgttgtggcc
     1861 tgatgctgga taactggaaa ctaggagaca gtcctattca aaagcataaa cagctatatc
     1921 ctagctgtag ctttattcag aatctggttt cagctagtct gggatccacc tctaagaata
     1981 cgtctccaat gagaaacagt tttgcacatt cattatctcc caccttggaa catagtagct
     2041 tgttcagtgg ttcttactcc agcctttctc caaaccctct taattctaga gcagttgaag
     2101 acatctcttc atcgaggact aacccctaca gttatgcaat gagtactgaa gaagccagat
     2161 ttcttaccta ccatatgtgg ccattaactt ttttgtcacc atcagaattg gcaagagctg
     2221 gtttttatta tataggacct ggagataggg tagcctgctt tgcctgtggt gggaagctca
     2281 gtaactggga accaaaggat gatgctatgt cagaacaccg gaggcatttt cccaactgtc
     2341 catttttgga aaattctcta gaaactctga ggtttagcat ttcaaatctg agcatgcaga
     2401 cacatgcagc tcgaatgaga acatttatgt actggccatc tagtgttcca gttcagcctg
     2461 agcagcttgc aagtgctggc ttttattatg tgggtcgcaa tgatgatgtc aaatgctttt
     2521 gttgtgatgg tggcttgagg tgttgggaat ctggagatga tccatgggta gaacatgcca
     2581 agtggtttcc aaggtgtgag ttcttgatac gaatgaaagg ccaagagttt gttgatgaga
     2641 ttcaaggtag atatcctcat cttcttgaac agctgttgtc aacttcagat accactggag
     2701 aagaaaatgc tgacccacca attattcatt ttggacctgg agaaagttct tcagaagatg
     2761 ctgtcatgat gaatacacct gtggttaaat ctgccttgga aatgggcttt aatagagacc
     2821 tggtgaaaca aacagttcaa agtaaaatcc tgacaactgg agagaactat aaaacagtta
     2881 atgatattgt gtcagcactt cttaatgctg aagatgaaaa aagagaagag gagaaggaaa
     2941 aacaagctga agaaatggca tcagatgatt tgtcattaat tcggaagaac agaatggctc
     3001 tctttcaaca attgacatgt gtgcttccta tcctggataa tcttttaaag gccaatgtaa
     3061 ttaataaaca ggaacatgat attattaaac aaaaaacaca gataccttta caagcgagag
     3121 aactgattga taccattttg gttaaaggaa atgctgcggc caacatcttc aaaaactgtc
     3181 taaaagaaat tgactctaca ttgtataaga acttatttgt ggataagaat atgaagtata
     3241 ttccaacaga agatgtttca ggtctgtcac tggaagaaca attgaggagg ttgcaagaag
     3301 aacgaacttg taaagtgtgt atggacaaag aagtttctgt tgtatttatt ccttgtggtc
     3361 atctggtagt atgccaggaa tgtgcccctt ctctaagaaa atgccctatt tgcaggggta
     3421 taatcaaggg tactgttcgt acatttctct ctgtttaaac gaattcgggc tcggtacccg
     3481 gggatcctct agagtcgacc tgcaggcatg caagctgatc cggctgctaa caaagcccga
     3541 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcg
     3601 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3661 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3721 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3781 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3841 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     3901 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     3961 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     4021 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4081 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4141 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4201 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4261 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4321 tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa tcatcggcat
     4381 agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag gcagatcatg
     4441 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4501 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4561 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4621 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4681 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4741 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4801 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4861 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4921 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4981 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5041 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5101 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5161 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5221 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5281 catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     5341 tgtgaatcga tagcgataag gatccgctca tgcgatgata agctgtcaaa catgagaatt
     5401 acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata aattgcactg
     5461 aaatctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc
     5521 gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc
     5581 tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac
     5641 ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa
     5701 ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga
     5761 tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc
     5821 ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca
     5881 taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc
     5941 gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     6001 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
     6061 ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt
     6121 tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
     6181 gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc
     6241 ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac
     6301 actccgctag cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga
     6361 ttacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     6421 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     6481 gagatct
//
