LOCUS       sequence-391338-        6134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human RNAP1 promoter region driving firefly luciferase expression 
            from IRES. Constructed by Ghoshal, et al. J Biol Chem (2004). Please
            cite this original manuscript!.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6134)
  AUTHORS   McCool MA, Buhagiar AF, Bryant CJ, Ogawa LM, Abriola L, Surovtseva 
            YV, Baserga SJ
  TITLE     Human pre-60S assembly factors link rRNA transcription to pre-rRNA 
            processing.
  JOURNAL   RNA. 2022 Nov 2. pii: rna.079149.122. doi: 10.1261/rna.079149.122.
  PUBMED    36323459
REFERENCE   2  (bases 1 to 6134)
  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: "RNA. 2022 
            Nov 2. pii: rna.079149.122. doi: 10.1261/rna.079149.122."
COMMENT     Sequence Label: pHrD-IRES-Fluc
FEATURES             Location/Qualifiers
     source          1..6134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    835..1405
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(999..1016)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1226..1245
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1419..3071
                     /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(1485..1503)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3112..3233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3123..3142)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3177..3196
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3481..3498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3652..4240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3732..3751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4411..5202,5203..5271))
                     /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     5034..5053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5272..5376)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5403..5858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5490..5509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5700..5721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5989..6037
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6051..6134,1..8)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6091..6110
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atttctctat cgataggtac cgagctctcc catatggtcg acctgcaggc ggccgcgaat
       61 tcactagtga ttaccccgcg atcctttctg gagagtcccc gtgcggagtc ggagagcgct
      121 ccctgagcgc gcgtgcggcc cgagaggtcg cgcctggccg gccttcggtc cctcgtgtgt
      181 cccggtcgta ggaggggccg gccgaaaatg cttccggctc ccgctctgga gacacgggcc
      241 ggccccctgc gtgtggcacg ggcggccggg agggcgtccc cggcccggcg ctgctcccgc
      301 gtgtgtcctg gggttgacca gagggccccg ggcgctccgt gtgtggctgc gatggtggcg
      361 tttttgggga caggtgtccg tgtcgcgcgt cgcctgggcc ggcggcgtgg tcggtgacgc
      421 gacctcccgg ccccggggga ggtatatctt tcgctccgag tcggcatttt gggccgccgg
      481 gttattgctg acacgctgtc ctctggcgac ctgtcgctgg agaggttggg cctccggatg
      541 cgcgcggggc tctggcctac cggtgacccg gctagccggc cgcgctcctg cttgagccgc
      601 ctgccggggc ccgcgggcct gctgttctct cgcgcgtccg agcgtcccga ctcccggtgc
      661 cggcccgggt ccgggtctct gacccacccg ggggcggcgg ggaaggcggc gagggccacc
      721 gtgcccccgt gcgctctccg ctgcgggcgc ccggggcggc cgcgacaacc ccaccccgct
      781 ggctccgtgc cgtgcgtgtc aggcgttctc gtcagatcta gggcggccaa ttcgcccctc
      841 tccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
      901 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
      961 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     1021 tctgttgaat gtcgtgaagg aagcagtacc tctggaagct tcttgaagac aaacaacgtc
     1081 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     1141 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     1201 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     1261 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     1321 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     1381 tttcctttga aaaacacgat gataagcttg ccacaaccat ggaagacgcc aaaaacataa
     1441 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
     1501 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
     1561 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
     1621 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     1681 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     1741 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     1801 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     1861 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1921 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     1981 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     2041 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     2101 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     2161 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     2221 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     2281 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     2341 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     2401 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     2461 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     2521 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     2581 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     2641 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     2701 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     2761 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     2821 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2881 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2941 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     3001 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     3061 tcgccgtgta attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     3121 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3181 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3241 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     3301 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
     3361 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     3421 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     3481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3841 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3961 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4021 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4081 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4141 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4201 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4261 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4321 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4381 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4441 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4501 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4561 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4621 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4681 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4741 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4801 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4861 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4921 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4981 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5041 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5101 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5161 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5221 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5281 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5341 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5401 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5461 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5581 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5701 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5761 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5821 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     5881 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     5941 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     6001 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     6061 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     6121 gcaggtgcca gaac
//
