LOCUS       sequence-479814-        6027 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express SmBiT-SRPK1 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 6027)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: SmBiT-SRPK1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..753
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(2970..2988)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3025..3146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3036..3055)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3090..3109
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3391..3748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3391..3582
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3414..3434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3599..3734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3763..3810
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3842..4636
                     /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(3896..3915)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4506..4525
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4700..4748
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4987..5575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5067..5086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5691..5974
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 aaatgagtct tcggacctcg cgggggccgc ttaagcggtg gttagggttt gtctgacgcg
       61 gggggagggg gaaggaacga aacactctca ttcggaggcg gctcggggtt tggtcttggt
      121 ggccacgggc acgcagaaga gcgccgcgat cctcttaagc acccccccgc cctccgtgga
      181 ggcgggggtt tggtcggcgg gtggtaactg gcgggccgct gactcgggcg ggtcgcgcgc
      241 cccagagtgt gaccttttcg gtctgctcgc agacccccgg gcggcgccgc cgcggcggcg
      301 acgggctcgc tgggtcctag gctccatggg gaccgtatac gtggacaggc tctggagcat
      361 ccgcacgact gcggtgatat taccggagac cttctgcggg acgagccggg tcacgcggct
      421 gacgcggagc gtccgttggg cgacaaacac caggacgggg cacaggtaca ctatcttgtc
      481 acccggaggc gcgagggact gcaggagctt cagggagtgg cgcagctgct tcatccccgt
      541 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
      601 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaagtcga acacgcagat
      661 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
      721 gaacaccgag cgaccctgca gcgacccgct taaaagcttg gcaatccggt actgttggta
      781 aagccaccgc tagcgctcac catggtgacc ggctaccggc tgttcgagga gattctcggg
      841 agctccggtg gtggcgggag cggaggtgga ggctcgagcg gtgcgatcgc catggagcgg
      901 aaagtgcttg cgctccaggc ccgaaagaaa aggaccaagg ccaagaagga caaagcccaa
      961 aggaaatctg aaactcagca ccgaggctct gctccccact ctgagagtga tctaccagag
     1021 caggaagagg agattctggg atctgatgat gatgagcaag aagatcctaa tgattattgt
     1081 aaaggaggtt atcatcttgt gaaaattgga gatctattca atgggagata ccatgtgatc
     1141 cgaaagttag gctggggaca cttttcaaca gtatggttat catgggatat tcaggggaag
     1201 aaatttgtgg caatgaaagt agttaaaagt gctgaacatt acactgaaac agcactagat
     1261 gaaatccggt tgctgaagtc agttcgcaat tcagacccta atgatccaaa tagagaaatg
     1321 gttgttcaac tactagatga ctttaaaata tcaggagtta atggaacaca tatctgcatg
     1381 gtatttgaag ttttggggca tcatctgctc aagtggatca tcaaatccaa ttatcagggg
     1441 cttccactgc cttgtgtcaa aaaaattatt cagcaagtgt tacagggtct tgattattta
     1501 cataccaagt gccgtatcat ccacactgac attaaaccag agaacatctt attgtcagtg
     1561 aatgagcagt acattcggag gctggctgca gaagcaacag aatggcagcg atctggagct
     1621 cctccgcctt ccggatctgc agtcagtact gctccccagc ctaaaccagc tgacaaaatg
     1681 tcaaagaata agaagaagaa attgaagaag aagcagaagc gccaggcaga attactagag
     1741 aagcgaatgc aggaaattga ggaaatggag aaagagtcgg gccctgggca aaaaagacca
     1801 aacaagcaag aagaatcaga gagtcctgtt gaaagaccct tgaaagagaa cccacctaat
     1861 aaaatgaccc aagaaaaact tgaagagtca agtaccattg gccaggatca aacgcttatg
     1921 gaacgtgata cagagggtgg tgcagcagaa attaattgca atggagtgat tgaagtcatt
     1981 aattatactc agaacagtaa taatgaaaca ttgagacata aagaggatct acataatgct
     2041 aatgactgtg atgtccaaaa tttgaatcag gaatctagtt tcctaagctc ccaaaatgga
     2101 gacagcagca catctcaaga aacagactct tgtacaccta taacatctga ggtgtcagac
     2161 accatggtgt gccagtcttc ctcaactgta ggtcagtcat tcagtgaaca acacattagc
     2221 caacttcaag aaagcattcg ggcagagata ccctgtgaag atgaacaaga gcaagaacat
     2281 aacggaccac tggacaacaa aggaaaatcc acggctggaa attttcttgt taatcccctt
     2341 gagccaaaaa atgcagaaaa gctcaaggtg aagattgctg accttggaaa tgcttgttgg
     2401 gtgcacaaac atttcactga agatattcaa acaaggcaat atcgttcctt ggaagttcta
     2461 atcggatctg gctataatac ccctgctgac atttggagca cggcatgcat ggcctttgaa
     2521 ctggccacag gtgactattt gtttgaacct cattcagggg aagagtacac tcgagatgaa
     2581 gatcacattg cattgatcat agaacttctg gggaaggtgc ctcgcaagct cattgtggca
     2641 ggaaaatatt ccaaggaatt tttcaccaaa aaaggtgacc tgaaacatat cacgaagctg
     2701 aaaccttggg gcctttttga ggttctagtg gagaagtatg agtggtcgca ggaagaggca
     2761 gctggcttca cagatttctt actgcccatg ttggagctga tccctgagaa gagagccact
     2821 gccgccgagt gtctccggca cccttggctt aactccgttt aaacgaattc gggctcggta
     2881 cccggggatc ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc
     2941 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     3001 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3061 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3121 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3181 ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3241 tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     3301 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     3361 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     3421 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3481 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3601 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3661 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3721 cttttttgga ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg
     3781 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat
     3841 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3901 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3961 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4021 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4081 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4141 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4201 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4261 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4321 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4381 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4441 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4501 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4561 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4621 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4681 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     4741 tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat
     4801 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctaccc
     4861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5401 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     5581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac
     5701 aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc
     5761 agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt
     5821 taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg
     5881 tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt
     5941 aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga
     6001 tgcgtccggc gtagaggatc gagatct
//
