LOCUS       sequence-476604-        6708 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express SRPK1-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 6708)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6708)
  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: SRPK1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6708
                     /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             3051..3563
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3651..3669)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3706..3827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3717..3736)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3771..3790
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4072..4429
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4072..4263
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4095..4115)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4280..4415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4444..4491
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4523..5317
                     /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(4577..4596)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5187..5206
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5381..5429
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5668..6256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5748..5767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6372..6655
                     /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 cgccatggag cggaaagtgc
     1081 ttgcgctcca ggcccgaaag aaaaggacca aggccaagaa ggacaaagcc caaaggaaat
     1141 ctgaaactca gcaccgaggc tctgctcccc actctgagag tgatctacca gagcaggaag
     1201 aggagattct gggatctgat gatgatgagc aagaagatcc taatgattat tgtaaaggag
     1261 gttatcatct tgtgaaaatt ggagatctat tcaatgggag ataccatgtg atccgaaagt
     1321 taggctgggg acacttttca acagtatggt tatcatggga tattcagggg aagaaatttg
     1381 tggcaatgaa agtagttaaa agtgctgaac attacactga aacagcacta gatgaaatcc
     1441 ggttgctgaa gtcagttcgc aattcagacc ctaatgatcc aaatagagaa atggttgttc
     1501 aactactaga tgactttaaa atatcaggag ttaatggaac acatatctgc atggtatttg
     1561 aagttttggg gcatcatctg ctcaagtgga tcatcaaatc caattatcag gggcttccac
     1621 tgccttgtgt caaaaaaatt attcagcaag tgttacaggg tcttgattat ttacatacca
     1681 agtgccgtat catccacact gacattaaac cagagaacat cttattgtca gtgaatgagc
     1741 agtacattcg gaggctggct gcagaagcaa cagaatggca gcgatctgga gctcctccgc
     1801 cttccggatc tgcagtcagt actgctcccc agcctaaacc agctgacaaa atgtcaaaga
     1861 ataagaagaa gaaattgaag aagaagcaga agcgccaggc agaattacta gagaagcgaa
     1921 tgcaggaaat tgaggaaatg gagaaagagt cgggccctgg gcaaaaaaga ccaaacaagc
     1981 aagaagaatc agagagtcct gttgaaagac ccttgaaaga gaacccacct aataaaatga
     2041 cccaagaaaa acttgaagag tcaagtacca ttggccagga tcaaacgctt atggaacgtg
     2101 atacagaggg tggtgcagca gaaattaatt gcaatggagt gattgaagtc attaattata
     2161 ctcagaacag taataatgaa acattgagac ataaagagga tctacataat gctaatgact
     2221 gtgatgtcca aaatttgaat caggaatcta gtttcctaag ctcccaaaat ggagacagca
     2281 gcacatctca agaaacagac tcttgtacac ctataacatc tgaggtgtca gacaccatgg
     2341 tgtgccagtc ttcctcaact gtaggtcagt cattcagtga acaacacatt agccaacttc
     2401 aagaaagcat tcgggcagag ataccctgtg aagatgaaca agagcaagaa cataacggac
     2461 cactggacaa caaaggaaaa tccacggctg gaaattttct tgttaatccc cttgagccaa
     2521 aaaatgcaga aaagctcaag gtgaagattg ctgaccttgg aaatgcttgt tgggtgcaca
     2581 aacatttcac tgaagatatt caaacaaggc aatatcgttc cttggaagtt ctaatcggat
     2641 ctggctataa tacccctgct gacatttgga gcacggcatg catggccttt gaactggcca
     2701 caggtgacta tttgtttgaa cctcattcag gggaagagta cactcgagat gaagatcaca
     2761 ttgcattgat catagaactt ctggggaagg tgcctcgcaa gctcattgtg gcaggaaaat
     2821 attccaagga atttttcacc aaaaaaggtg acctgaaaca tatcacgaag ctgaaacctt
     2881 ggggcctttt tgaggttcta gtggagaagt atgagtggtc gcaggaagag gcagctggct
     2941 tcacagattt cttactgccc atgttggagc tgatccctga gaagagagcc actgccgccg
     3001 agtgtctccg gcacccttgg cttaactccg tttctctcgg ctcgagcggc gtcttcacac
     3061 tcgaagattt cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg
     3121 aacagggagg tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa
     3181 ggattgtcct gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg
     3241 aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg
     3301 tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta
     3361 cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca
     3421 aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga
     3481 tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc
     3541 tgtgcgaacg cattctggcg taaggccgcg actctagagt cgacctgcag gcatgcaagc
     3601 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     3661 ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg
     3721 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3781 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3841 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa
     3901 acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga
     3961 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     4021 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     4081 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4261 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4321 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4381 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta
     4441 actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     4501 aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc
     4561 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4621 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4681 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4741 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4801 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4861 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4921 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4981 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5041 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5101 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5161 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5221 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5281 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5341 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     5401 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     5461 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     5521 ctgcggactg gctttctacc cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5581 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6181 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6241 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6361 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     6421 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     6481 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     6541 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     6601 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     6661 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
