LOCUS       sequence-217335-        5859 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 5859)
  AUTHORS   Petersen CP, Bordeleau ME, Pelletier J, Sharp PA
  TITLE     Short RNAs Repress Translation after Initiation in Mammalian Cells.
  JOURNAL   Mol Cell. 2006 Feb 17. 21(4):533-42.
  PUBMED    16483934
REFERENCE   2  (bases 1 to 5859)
  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: "Mol Cell. 
            2006 Feb 17. 21(4):533-42."
COMMENT     Sequence Label: pFR_CrPV_xb
FEATURES             Location/Qualifiers
     source          1..5859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        664..1415
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      1445..1454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1451..3103
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1517..1535)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             3336..4271
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    4470..4591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4481..4500)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4535..4554
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4724..4828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4829..4897,4898..5689)
                     /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(5047..5066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
      661 ctaaatgagt cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg
      721 cggggggagg gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg
      781 gtggccacgg gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg
      841 gaggcggggg tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc
      901 gccccagagt gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg
      961 cgacgggctc gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc
     1021 atccgcacga ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg
     1081 ctgacgcgga gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg
     1141 tcacccggag gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc
     1201 gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt
     1261 agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag
     1321 atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc
     1381 tcgaacaccg agcgaccctg cagcgacccg cttaaaagct tggcattccg gtactgttgg
     1441 taaagccacc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
     1501 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     1561 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     1621 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     1681 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     1741 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     1801 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     1861 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     1921 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     1981 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     2041 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     2101 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     2161 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     2221 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct
     2281 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     2341 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     2401 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     2461 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     2521 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     2581 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     2641 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     2701 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg
     2761 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     2821 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc
     2881 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     2941 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     3001 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     3061 gatcctcata aaggccaaga agggcggaaa gatcgccgtg taattctagc actagtaaag
     3121 caaaaatgtg atcttgcttg taaatacaat tttgagaggt taataaatta caagtagtgc
     3181 tatttttgta tttaggttag ctatttagct ttacgttcca ggatgcctag tggcagcccc
     3241 acaatatcca ggaagccctc tctgcggttt ttcagattag gtagtcgaaa aacctaagaa
     3301 atttacctgc tacatttcaa gataatctag ccaccatgac ttcgaaagtt tatgatccag
     3361 aacaaaggaa acggatgata actggtccgc agtggtgggc cagatgtaaa caaatgaatg
     3421 ttcttgattc atttattaat tattatgatt cagaaaaaca tgcagaaaat gctgttattt
     3481 ttttacatgg taacgcggcc tcttcttatt tatggcgaca tgttgtgcca catattgagc
     3541 cagtagcgcg gtgtattata ccagacctta ttggtatggg caaatcaggc aaatctggta
     3601 atggttctta taggttactt gatcattaca aatatcttac tgcatggttt gaacttctta
     3661 atttaccaaa gaagatcatt tttgtcggcc atgattgggg tgcttgtttg gcatttcatt
     3721 atagctatga gcatcaagat aagatcaaag caatagttca cgctgaaagt gtagtagatg
     3781 tgattgaatc atgggatgaa tggcctgata ttgaagaaga tattgcgttg atcaaatctg
     3841 aagaaggaga aaaaatggtt ttggagaata acttcttcgt ggaaaccatg ttgccatcaa
     3901 aaatcatgag aaagttagaa ccagaagaat ttgcagcata tcttgaacca ttcaaagaga
     3961 aaggtgaagt tcgtcgtcca acattatcat ggcctcgtga aatcccgtta gtaaaaggtg
     4021 gtaaacctga cgttgtacaa attgttagga attataatgc ttatctacgt gcaagtgatg
     4081 atttaccaaa aatgtttatt gaatcggacc caggattctt ttccaatgct attgttgaag
     4141 gtgccaagaa gtttcctaat actgaatttg tcaaagtaaa aggtcttcat ttttcgcaag
     4201 aagatgcacc tgatgaaatg ggaaaatata tcaaatcgtt cgttgagcga gttctcaaaa
     4261 atgaacaata attctagact cgagccggaa gttttcacaa agctaacacc ggaagttttc
     4321 acaaagctaa caccggaagt tttcacaaag ctaacaccgg aagttttcac aaagctaaca
     4381 ccggagtttt cacaaagcta acaccggaag ttttcacaaa gctaacaccg gatcgcgggc
     4441 cctagagcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4501 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4561 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4621 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4681 taaaatcgat aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4801 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4861 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4921 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4981 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5041 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5101 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5161 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5221 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5281 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5341 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5401 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5461 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5521 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5581 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5641 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5701 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
