LOCUS       sequence-12564-e        4183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4183)
  AUTHORS   Trujillo RD, Yue SB, Tang Y, O'Gorman WE, Chen CZ
  TITLE     The potential functions of primary microRNAs in target recognition 
            and repression.
  JOURNAL   EMBO J. 2010 Oct 6;29(19):3272-85. doi: 10.1038/emboj.2010.208. Epub
            2010 Aug 31.
  PUBMED    20808284
REFERENCE   2  (bases 1 to 4183)
  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"; doi: 
            "10.1038/emboj.2010.208"; journalName: "EMBO J"; date: "2010-10-6- 
            6"; volume: "29"; issue: "19"; pages: "3272-85"
COMMENT     Sequence Label: phRL-TKlk-lin-41_LCS-2G
FEATURES             Location/Qualifiers
     source          1..4183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..759
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          829..961
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     944..963
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1006..1024
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1028..1037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1034..1969
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2138..2259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2149..2168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2203..2222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2392..2496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2497..2565,2566..3357)
                     /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(2715..2734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3528..4116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4017..4036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
       61 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
      121 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
      181 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
      241 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
      301 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
      361 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
      421 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
      481 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
      541 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
      601 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
      661 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
      721 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc ttctgacaca
      781 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      841 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      901 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
      961 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
     1021 taggctagcc accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac
     1081 tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta
     1141 ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc
     1201 cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc
     1261 tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga
     1321 tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt
     1381 tgtgggccac gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa
     1441 gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg
     1501 gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct
     1561 tgagaataac ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc
     1621 tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac
     1681 cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat
     1741 tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga
     1801 gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac
     1861 cgagttcgtg aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg
     1921 taagtacatc aagagcttcg tggagcgcgt gctgaagaac gagcagtaat tctagaaccg
     1981 gtctcgagaa ctcaagtata ccttttatac aaccgttcta cactgaacgc gatgtaaata
     2041 tcgcaatccc tttttataca accattctgc ctgtgaacca ttgaaacctt gagctccata
     2101 tgggtacctc tagagcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2161 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2221 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2281 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2341 aaatgtggta aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc
     2401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2461 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2521 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2581 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2641 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2701 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2761 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2821 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2881 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2941 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3001 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3061 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3121 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3181 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3241 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3301 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3361 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3421 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3481 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3541 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3601 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3661 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3721 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3781 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3841 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3901 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3961 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4021 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4081 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4141 cggttcctgg ccttttgctg gccttttgct cacatggctc gac
//
