LOCUS       Exported                5843 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Tagged-IRES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  AUTHORS   Tsai BP, Wang X, Huang L, Waterman ML
  TITLE     Quantitative profiling of in vivo-assembled RNA-protein complexes 
            using a novel integrated proteomic approach.
  JOURNAL   Mol Cell Proteomics. 2011 Apr;10(4):M110.007385. Epub 2011 Feb 1.
  PUBMED    21285413
REFERENCE   2  (bases 1 to 5843)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5843
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        66..423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(89..109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      274..409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     336..355
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          493..625
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     608..627
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        670..688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1927..3579
                     /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(1993..2011)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_RNA        3631..3649
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3667..3685
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3703..3721
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     polyA_signal    3790..3911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3801..3820)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3855..3874
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4052..4156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4157..5017
                     /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(4375..4394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5188..5776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5677..5696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcgatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
       61 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      421 aaaaagcttg attcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga
      481 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      541 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      601 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      661 tagagtactt aatacgactc actataggct agaattcggt accccgggtt cgaaatcgat
      721 aagcttggat ctagcgcgcg ggaggaggag aagcagtggg gaggcgcagc cgctcacctg
      781 cggggcaggg cgcggaggag ggacccgggc tgcgcgctct cgggccgagg aaccaggacg
      841 cgcccggagc ctcgcacgcg gccaagctcg gggcgtcccc tcccctcggc cgggcgaact
      901 caaggggcgc agctctttgc tttgacagag ctggccggcg gaggcgtgca gagcggcgag
      961 ccggcgagcc aggctgagaa actcgagccg ggaacaaaga ggggtcggac tgagtgtgtg
     1021 tgtcggctcg agctccgggc agaggcattt gggcccgagg cccccgctgt gactccccga
     1081 gactccgcag tgccctccac tgcggagtcc ccgcgcttgc cggcaaaaac tttattcttg
     1141 gcaaacttct ctttctcttc ccctcctcct cggcccccat cttctgctcc tcctccttct
     1201 ctagcagatt aaatgagcct cgagaagaaa aaccgaagcg aaagggaaga aaataagaag
     1261 atctaaaacg gacatctcca gcgtgggtgg ctcctttttc tttttctttt tttcccaccc
     1321 ttcaggaagt ggacgtttcg ttatcttctg atccttgcac cttcttttgg ggcaaacggg
     1381 gcccttctgc ccagatcccc tctcttttct cggaaaacaa actactaagt cggcatccgg
     1441 ggtaactaca gtggagaggg tttccgcgga gacgcgccgc cggaccctcc tctgcacttt
     1501 ggggaggcgt gctccctcca gaaccggcgt tctccgcgcg caaatcccgg cgacgcgggg
     1561 tcgcggggtg gccgccgggg cagcctcgtc tagcgcgcgc cgcgcagacg cccccggagt
     1621 cgccagctac cgcagccctc gccgcccagt gcccttcggc ctcgggggcg ggcgcctgcg
     1681 tcggtctccg cgaagcggga aagcgcggcg gccgccggga ttcgggcgcc gcggcagctg
     1741 ctccggctgc cggccggcgg ccccgcgctc gcccgccccg cttccgcccg ctgtcctgct
     1801 gcacgaaccc ttccaactct cctttcctcc cccacccttg agttacccct ctgtctttcc
     1861 tgctgttgcg cgggtgctcc cacagcggag cggagattac agagccgccg ggatgcccct
     1921 ggatccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa
     1981 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga
     2041 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa
     2101 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc
     2161 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc
     2221 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg
     2281 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac
     2341 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac
     2401 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa
     2461 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc
     2521 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga
     2581 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta
     2641 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt
     2701 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag
     2761 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc
     2821 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc
     2881 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa
     2941 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa
     3001 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc
     3061 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg
     3121 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta
     3181 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg
     3241 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg
     3301 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt
     3361 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc
     3421 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt
     3481 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc
     3541 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat tctagatcat cgaaacatga
     3601 ggatcaccca tatctgcagt cgacatcgaa acatgaggat cacccatgtc tgcagtcgac
     3661 atcgaaacat gaggatcacc catgtctgca gtcgacatcg aaacatgagg atcacccatg
     3721 tctgcagtcg acatcgaaat cgataagctt cagatcagat cctagagcgg ccgcttcgag
     3781 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3841 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3901 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     3961 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aagtttaaac
     4021 agatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4141 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4201 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4261 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4321 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4381 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4441 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4501 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4561 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4621 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4681 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4741 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4801 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4861 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4921 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4981 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5821 gccttttgct cacatggctc gac
//
