LOCUS       sequence-117540-        6066 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 6066)
  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 6066)
  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_HCV_xb
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..759
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      789..798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             795..2447
                     /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(861..879)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2887..3822
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    4021..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4032..4051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4086..4105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4275..4379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4380..4448,4449..5240)
                     /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(4598..4617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5411..5999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5900..5919
                     /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 agcttggcat tccggtactg
      781 ttggtaaagc caccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
      841 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
      901 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
      961 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     1021 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     1081 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     1141 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     1201 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     1261 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     1321 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     1381 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     1441 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     1501 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     1561 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     1621 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     1681 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     1741 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     1801 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     1861 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     1921 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     1981 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     2041 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     2101 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     2161 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     2221 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     2281 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     2341 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     2401 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagtgtacgt
     2461 agccagcccc cgattggggg cgacactcca ccatagatca ctcccctgtg aggaactact
     2521 gtcttcacgc agaaagcgtc tagccatggc gttagtatga gagtcgtgca gcctccagga
     2581 ccccccctcc cgggagagcc atagtggtct gcggaaccgg tgagtacacc ggaattgcca
     2641 ggacgaccgg gtcctttctt ggatcaaccc gctcaatgcc tggagatttg ggcgtgcccc
     2701 cgcaagactg ctagccgagt agtgttgggt cgcgaaaggc cttgtggtac tgcctgatag
     2761 ggtgcttgcg agtgccccgg gaggtctcgt agaccgtgca ccatgagcac gaatcctaaa
     2821 cctcaaagaa aaaccaaacg taacaccaac cgccgcccac aggacgtctt catatgtcta
     2881 gccaccatga cttcgaaagt ttatgatcca gaacaaagga aacggatgat aactggtccg
     2941 cagtggtggg ccagatgtaa acaaatgaat gttcttgatt catttattaa ttattatgat
     3001 tcagaaaaac atgcagaaaa tgctgttatt tttttacatg gtaacgcggc ctcttcttat
     3061 ttatggcgac atgttgtgcc acatattgag ccagtagcgc ggtgtattat accagacctt
     3121 attggtatgg gcaaatcagg caaatctggt aatggttctt ataggttact tgatcattac
     3181 aaatatctta ctgcatggtt tgaacttctt aatttaccaa agaagatcat ttttgtcggc
     3241 catgattggg gtgcttgttt ggcatttcat tatagctatg agcatcaaga taagatcaaa
     3301 gcaatagttc acgctgaaag tgtagtagat gtgattgaat catgggatga atggcctgat
     3361 attgaagaag atattgcgtt gatcaaatct gaagaaggag aaaaaatggt tttggagaat
     3421 aacttcttcg tggaaaccat gttgccatca aaaatcatga gaaagttaga accagaagaa
     3481 tttgcagcat atcttgaacc attcaaagag aaaggtgaag ttcgtcgtcc aacattatca
     3541 tggcctcgtg aaatcccgtt agtaaaaggt ggtaaacctg acgttgtaca aattgttagg
     3601 aattataatg cttatctacg tgcaagtgat gatttaccaa aaatgtttat tgaatcggac
     3661 ccaggattct tttccaatgc tattgttgaa ggtgccaaga agtttcctaa tactgaattt
     3721 gtcaaagtaa aaggtcttca tttttcgcaa gaagatgcac ctgatgaaat gggaaaatat
     3781 atcaaatcgt tcgttgagcg agttctcaaa aatgaacaat aattctagac tcgagccgga
     3841 agttttcaca aagctaacac cggaagtttt cacaaagcta acaccggaag ttttcacaaa
     3901 gctaacaccg gaagttttca caaagctaac accggagttt tcacaaagct aacaccggaa
     3961 gttttcacaa agctaacacc ggatcgcggg ccctagagcg gccgcttcga gcagacatga
     4021 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4081 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4141 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4201 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatcca ggtggcactt
     4261 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4381 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4441 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4501 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4561 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4621 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4681 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4741 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4801 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4861 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4921 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4981 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5041 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5101 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5161 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5221 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5281 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5341 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5521 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5761 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5821 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5881 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5941 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6001 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     6061 ctcgac
//
