LOCUS       sequence-351976-        6521 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 6521)
  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 6521)
  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 Cdk2ap1Exon1
FEATURES             Location/Qualifiers
     source          1..6521
                     /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"
     primer_bind     666..685
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             942..1877
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1936..1984
                     /note="synthetic polyadenylation signal"
     promoter        1993..2744
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2774..2783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2780..4432
                     /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             3739..3747
                     /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    4476..4597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4487..4506)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4541..4560
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4730..4834
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4835..4903,4904..5695)
                     /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(5053..5072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5866..6454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6355..6374
                     /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 tagggcggcc gcccctgcgc cgcgcgcccg atgtatgggt
      721 gatctactgc ggcggcggcg gcggcggcgg cggcagggtg agggacggcc atatttgccg
      781 gcgcggcccg agccggcgac aacaaaaagt gcgcgggctc ccggcgggcg ctcggacggc
      841 tgcggggacg tggggccgcc cggcccgggc ctcggcctcg gcctcgccgg cgcgctcgcg
      901 gcctggcccg gctctgcccg gcgcgcccgc cgcccgcggg gatggcttcc aaggtgtacg
      961 accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa
     1021 tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc gagaacgccg
     1081 tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca
     1141 tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag tccggcaaga
     1201 gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct tggttcgagc
     1261 tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct tgtctggcct
     1321 ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct gagagtgtcg
     1381 tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc gccctgatca
     1441 agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag accatgctcc
     1501 caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg gagccattca
     1561 aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc cctctcgtta
     1621 agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac cttcgggcca
     1681 gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc aacgctattg
     1741 tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc ctccacttca
     1801 gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc
     1861 tgaagaacga gcagtaattc taggcgatcg ctcgagcccg ggaattcgtt taaacctaga
     1921 gcggccgctg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     1981 tgtgaggatc taaatgagtc ttcggacctc gcgggggccg cttaagcggt ggttagggtt
     2041 tgtctgacgc ggggggaggg ggaaggaacg aaacactctc attcggaggc ggctcggggt
     2101 ttggtcttgg tggccacggg cacgcagaag agcgccgcga tcctcttaag cacccccccg
     2161 ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact ggcgggccgc tgactcgggc
     2221 gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg cagacccccg ggcggcgccg
     2281 ccgcggcggc gacgggctcg ctgggtccta ggctccatgg ggaccgtata cgtggacagg
     2341 ctctggagca tccgcacgac tgcggtgata ttaccggaga ccttctgcgg gacgagccgg
     2401 gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca ccaggacggg gcacaggtac
     2461 actatcttgt cacccggagg cgcgagggac tgcaggagct tcagggagtg gcgcagctgc
     2521 ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg
     2581 catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg
     2641 aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat taaggtgacg
     2701 cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaaaagctt ggcattccgg
     2761 tactgttggt aaagccacca tggccgatgc taagaacatt aagaagggcc ctgctccctt
     2821 ctaccctctg gaggatggca ccgctggcga gcagctgcac aaggccatga agaggtatgc
     2881 cctggtgcct ggcaccattg ccttcaccga tgcccacatt gaggtggaca tcacctatgc
     2941 cgagtacttc gagatgtctg tgcgcctggc cgaggccatg aagaggtacg gcctgaacac
     3001 caaccaccgc atcgtggtgt gctctgagaa ctctctgcag ttcttcatgc cagtgctggg
     3061 cgccctgttc atcggagtgg ccgtggcccc tgctaacgac atttacaacg agcgcgagct
     3121 gctgaacagc atgggcattt ctcagcctac cgtggtgttc gtgtctaaga agggcctgca
     3181 gaagatcctg aacgtgcaga agaagctgcc tatcatccag aagatcatca tcatggactc
     3241 taagaccgac taccagggct tccagagcat gtacacattc gtgacatctc atctgcctcc
     3301 tggcttcaac gagtacgact tcgtgccaga gtctttcgac agggacaaaa ccattgccct
     3361 gatcatgaac agctctgggt ctaccggcct gcctaagggc gtggccctgc ctcatcgcac
     3421 cgcctgtgtg cgcttctctc acgcccgcga ccctattttc ggcaaccaga tcatccccga
     3481 caccgctatt ctgagcgtgg tgccattcca ccacggcttc ggcatgttca ccaccctggg
     3541 ctacctgatt tgcggctttc gggtggtgct gatgtaccgc ttcgaggagg agctgttcct
     3601 gcgcagcctg caagactaca aaattcagtc tgccctgctg gtgccaaccc tgttcagctt
     3661 cttcgctaag agcaccctga tcgacaagta cgacctgtct aacctgcacg agattgcctc
     3721 tggcggcgcc ccactgtcta aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc
     3781 aggcatccgc cagggctacg gcctgaccga gacaaccagc gccattctga ttaccccaga
     3841 gggcgacgac aagcctggcg ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt
     3901 ggacctggac accggcaaga ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg
     3961 ccctatgatt atgtccggct acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa
     4021 ggacggctgg ctgcactctg gcgacattgc ctactgggac gaggacgagc acttcttcat
     4081 cgtggaccgc ctgaagtctc tgatcaagta caagggctac caggtggccc cagccgagct
     4141 ggagtctatc ctgctgcagc accctaacat tttcgacgcc ggagtggccg gcctgcccga
     4201 cgacgatgcc ggcgagctgc ctgccgccgt cgtcgtgctg gaacacggca agaccatgac
     4261 cgagaaggag atcgtggact atgtggccag ccaggtgaca accgccaaga agctgcgcgg
     4321 cggagtggtg ttcgtggacg aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa
     4381 gatccgcgag atcctgatca aggctaagaa aggcggcaag atcgccgtgt aataattcta
     4441 gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     4501 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4561 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4621 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     4681 atgtggtaaa atcgataagg atccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4741 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4801 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4861 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4921 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4981 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5041 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5101 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5161 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5221 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5281 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5341 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5401 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5461 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5521 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5581 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5641 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5701 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5761 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5821 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5881 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5941 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6001 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6061 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6121 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6181 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6241 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6301 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6361 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6421 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6481 gttcctggcc ttttgctggc cttttgctca catggctcga c
//
