LOCUS       sequence-351978-        6398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression vector to test translation efficiency by luciferase 
            reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  AUTHORS   Modzelewski AJ, Shao W, Chen J, Lee A, Qi X, Noon M, Tjokro K, Sales
            G, Biton A, Anand A, Speed TP, Xuan Z, Wang T, Risso D, He L
  TITLE     A mouse-specific retrotransposon drives a conserved Cdk2ap1 isoform 
            essential for development.
  JOURNAL   Cell. 2021 Oct 7. pii: S0092-8674(21)01104-1. doi: 
            10.1016/j.cell.2021.09.021.
  PUBMED    34644528
REFERENCE   2  (bases 1 to 6398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2021 
            Oct 7. pii: S0092-8674(21)01104-1. doi: 10.1016/j.cell.2021.09.021."
COMMENT     Sequence Label: psiCheck Cdk2ap1-Delta-N (MT2B2)
FEATURES             Location/Qualifiers
     source          1..6398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             819..1754
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1813..1861
                     /note="synthetic polyadenylation signal"
     promoter        1870..2621
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2651..2660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2657..4309
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3616..3624
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4353..4474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4364..4383)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4418..4437
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4607..4711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4712..4780,4781..5572)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4930..4949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5743..6331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6232..6251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggacacat acacttcttt gcctgcttgc cttgtgggac
      721 tgaacaactg ctagatcctt ggacttccat tcacagctgc tgctgaccgt tgtcggagtt
      781 ggactgcaga cggaagtgtc cactcaccct ctactagcat ggcttccaag gtgtacgacc
      841 ccgagcaacg caaacgcatg atcactgggc ctcagtggtg ggctcgctgc aagcaaatga
      901 acgtgctgga ctccttcatc aactactatg attccgagaa gcacgccgag aacgccgtga
      961 tttttctgca tggtaacgct gcctccagct acctgtggag gcacgtcgtg cctcacatcg
     1021 agcccgtggc tagatgcatc atccctgatc tgatcggaat gggtaagtcc ggcaagagcg
     1081 ggaatggctc atatcgcctc ctggatcact acaagtacct caccgcttgg ttcgagctgc
     1141 tgaaccttcc aaagaaaatc atctttgtgg gccacgactg gggggcttgt ctggcctttc
     1201 actactccta cgagcaccaa gacaagatca aggccatcgt ccatgctgag agtgtcgtgg
     1261 acgtgatcga gtcctgggac gagtggcctg acatcgagga ggatatcgcc ctgatcaaga
     1321 gcgaagaggg cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc atgctcccaa
     1381 gcaagatcat gcggaaactg gagcctgagg agttcgctgc ctacctggag ccattcaagg
     1441 agaagggcga ggttagacgg cctaccctct cctggcctcg cgagatccct ctcgttaagg
     1501 gaggcaagcc cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt cgggccagcg
     1561 acgatctgcc taagatgttc atcgagtccg accctgggtt cttttccaac gctattgtcg
     1621 agggagctaa gaagttccct aacaccgagt tcgtgaaggt gaagggcctc cacttcagcc
     1681 aggaggacgc tccagatgaa atgggtaagt acatcaagag cttcgtggag cgcgtgctga
     1741 agaacgagca gtaattctag gcgatcgctc gagcccggga attcgtttaa acctagagcg
     1801 gccgctggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     1861 gaggatctaa atgagtcttc ggacctcgcg ggggccgctt aagcggtggt tagggtttgt
     1921 ctgacgcggg gggaggggga aggaacgaaa cactctcatt cggaggcggc tcggggtttg
     1981 gtcttggtgg ccacgggcac gcagaagagc gccgcgatcc tcttaagcac ccccccgccc
     2041 tccgtggagg cgggggtttg gtcggcgggt ggtaactggc gggccgctga ctcgggcggg
     2101 tcgcgcgccc cagagtgtga ccttttcggt ctgctcgcag acccccgggc ggcgccgccg
     2161 cggcggcgac gggctcgctg ggtcctaggc tccatgggga ccgtatacgt ggacaggctc
     2221 tggagcatcc gcacgactgc ggtgatatta ccggagacct tctgcgggac gagccgggtc
     2281 acgcggctga cgcggagcgt ccgttgggcg acaaacacca ggacggggca caggtacact
     2341 atcttgtcac ccggaggcgc gagggactgc aggagcttca gggagtggcg cagctgcttc
     2401 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     2461 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     2521 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     2581 gtggcctcga acaccgagcg accctgcagc gacccgctta aaagcttggc attccggtac
     2641 tgttggtaaa gccaccatgg ccgatgctaa gaacattaag aagggccctg ctcccttcta
     2701 ccctctggag gatggcaccg ctggcgagca gctgcacaag gccatgaaga ggtatgccct
     2761 ggtgcctggc accattgcct tcaccgatgc ccacattgag gtggacatca cctatgccga
     2821 gtacttcgag atgtctgtgc gcctggccga ggccatgaag aggtacggcc tgaacaccaa
     2881 ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc ttcatgccag tgctgggcgc
     2941 cctgttcatc ggagtggccg tggcccctgc taacgacatt tacaacgagc gcgagctgct
     3001 gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg tctaagaagg gcctgcagaa
     3061 gatcctgaac gtgcagaaga agctgcctat catccagaag atcatcatca tggactctaa
     3121 gaccgactac cagggcttcc agagcatgta cacattcgtg acatctcatc tgcctcctgg
     3181 cttcaacgag tacgacttcg tgccagagtc tttcgacagg gacaaaacca ttgccctgat
     3241 catgaacagc tctgggtcta ccggcctgcc taagggcgtg gccctgcctc atcgcaccgc
     3301 ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc aaccagatca tccccgacac
     3361 cgctattctg agcgtggtgc cattccacca cggcttcggc atgttcacca ccctgggcta
     3421 cctgatttgc ggctttcggg tggtgctgat gtaccgcttc gaggaggagc tgttcctgcg
     3481 cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg ccaaccctgt tcagcttctt
     3541 cgctaagagc accctgatcg acaagtacga cctgtctaac ctgcacgaga ttgcctctgg
     3601 cggcgcccca ctgtctaagg aggtgggcga agccgtggcc aagcgctttc atctgccagg
     3661 catccgccag ggctacggcc tgaccgagac aaccagcgcc attctgatta ccccagaggg
     3721 cgacgacaag cctggcgccg tgggcaaggt ggtgccattc ttcgaggcca aggtggtgga
     3781 cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc gagctgtgtg tgcgcggccc
     3841 tatgattatg tccggctacg tgaataaccc tgaggccaca aacgccctga tcgacaagga
     3901 cggctggctg cactctggcg acattgccta ctgggacgag gacgagcact tcttcatcgt
     3961 ggaccgcctg aagtctctga tcaagtacaa gggctaccag gtggccccag ccgagctgga
     4021 gtctatcctg ctgcagcacc ctaacatttt cgacgccgga gtggccggcc tgcccgacga
     4081 cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa cacggcaaga ccatgaccga
     4141 gaaggagatc gtggactatg tggccagcca ggtgacaacc gccaagaagc tgcgcggcgg
     4201 agtggtgttc gtggacgagg tgcccaaggg cctgaccggc aagctggacg cccgcaagat
     4261 ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc gccgtgtaat aattctagag
     4321 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     4381 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4441 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4501 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4561 tggtaaaatc gataaggatc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4621 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4681 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4741 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4801 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4861 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4921 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4981 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5041 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5101 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5161 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5221 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5281 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5341 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5401 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5461 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5521 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5581 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5881 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6361 cctggccttt tgctggcctt ttgctcacat ggctcgac
//
