LOCUS       MUT_S1-spacer_H1        6130 bp DNA     circular SYN 03-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6130)
  AUTHORS   Mauricio
  TITLE     Direct Submission
  JOURNAL   Exported Jul 15, 2023 from SnapGene Viewer 7.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6130
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     protein_bind    19..289
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        322..360
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     exon            375..535
                     /label=Exon 1
                     /note="The 5' end of this exon has not been accurately 
                     mapped. This exon is a hybrid between CMV IE1 exon 1 and 
                     CHO DHFR exon 1."
     misc_feature    392..416
                     /label=oMA179
     intron          536..834
                     /label=Intron 1
                     /note="This intron is derived from CHD DHFR intron 1. A Not
                     I site has been added to facilitate cloning procedures."
     misc_feature    835..1088
                     /label=Designer Exon
     misc_feature    836..1083
                     /label=SRSF1 binding region (15 modules)
     misc_feature    1086..1094
                     /label=Splice site (Major)
                     /note="Major spliceosome: consensus sequence CAGGTAAGT"
     intron          1089..2835
                     /label=intron 2
                     /note="This intron is derived from CHO DHFR intron 3 and 
                     also contains designed modules. It has the stalling 
                     mutation CAG -> CGG in the acceptor site."
     misc_feature    1095..1814
                     /label=spacer
                     /note="Reference modules"
     misc_feature    1815..2206
                     /label=hnRNP A1 binding region (24 modules)
     misc_feature    2546..2568
                     /label=cryptic 3'ss (MaxEnt: 9.4)
     misc_feature    2604..2626
                     /label=cryptic 3'ss (MaxEnt: 6.6)
     misc_feature    2647..2669
                     /label=cryptic 3'ss (MaxEnt: 2.8)
     misc_feature    2718..2740
                     /label=cryptic 3'ss (MaxEnt: 2.8)
     exon            2836..3608
                     /label=Exon 3
                     /note="This exon is derived from a concatenation of CHO 
                     DHFR exons 4, 5 and 6. The 3' end of this exon has not been
                     mapped accurately."
     primer_bind     complement(2917..2947)
                     /label=oMA203_Common3-R
     primer_bind     complement(2981..3001)
                     /label=cDNAGenerator
     misc_feature    3236..3258
                     /label=cryptic 3'ss (MaxEnt: 6.5)
     misc_feature    3260..3282
                     /label=cryptic 3'ss (MaxEnt: 6.2)
     misc_feature    3301..3323
                     /label=cryptic 3'ss (MaxEnt: 4.0)
     misc_feature    3420..3442
                     /label=cryptic 3'ss (MaxEnt: 5.2)
     polyA_signal    3460..3594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3465..3498
                     /label=oMA180
     misc_feature    3499..3516
                     /label=oMA178
     rep_origin      complement(4311..4899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5070..5930)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5931..6035)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttcactcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
       61 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
      121 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      181 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      241 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      301 cgagctcggt acccgggtcg aggtaggcgt gtacggtggg aggcctatat aagcagagct
      361 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      421 agacaccggg accgatccag cctccgcgta cggttcgacc gctgaactgc atcgtcgccg
      481 tgtcccagaa taagggcatc ggcaagaacg gagaccttcc ctggccaaag ctcaggtact
      541 ggctggattg ggttagggaa accgaggcgg ttcgctgaat cgggtcgagc acttggcgga
      601 gacgcgcggg ccaactactt agggacagtc atgaggggta ggcccgccgg ctgcagccct
      661 tgcccatgcc cgcggtgatc cccatgctgt gccagccttt gcccagaggc gctctagctg
      721 ggagcaaagt ccggtcactg ggcagcacca ccccccggac ttgcatgggt agccgctgag
      781 atggagcctg agcacacgcg gccgctgtta acgcagtgtt tctctttttt tcaggccaaa
      841 caagaggaac gccaaacaag aggaacgcca aacaagagga acgccaaaca agaggaacgc
      901 caaacaagag gaacgccaaa caagaggaac gccaaacaag aggaacgcca aacaagagga
      961 acgccaaaca agaggaacgc caaacaagag gaacgccaaa caagaggaac gccaaacaag
     1021 aggaacgcca aacaagagga acgccaaaca agaggaacgc caaacaagag gaacgccaaa
     1081 caacacaggt aagtccaaac aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac
     1141 aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc
     1201 aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac
     1261 aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc
     1321 aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac
     1381 aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc
     1441 aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac
     1501 aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc
     1561 aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac
     1621 aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc
     1681 aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac
     1741 aaccaaacaa ccaaacaacc aaacaaccaa acaaccaaac aaccaaacaa ccaaacaacc
     1801 aaacaaccaa acaaccaaac aattagggaa ccaaacaatt agggaaccaa acaattaggg
     1861 aaccaaacaa ttagggaacc aaacaattag ggaaccaaac aattagggaa ccaaacaatt
     1921 agggaaccaa acaattaggg aaccaaacaa ttagggaacc aaacaattag ggaaccaaac
     1981 aattagggaa ccaaacaatt agggaaccaa acaattaggg aaccaaacaa ttagggaacc
     2041 aaacaattag ggaaccaaac aattagggaa ccaaacaatt agggaaccaa acaattaggg
     2101 aaccaaacaa ttagggaacc aaacaattag ggaaccaaac aattagggaa ccaaacaatt
     2161 agggaaccaa acaattaggg aaccaaacaa ttagggaacc aaacaagcta gctggccagt
     2221 gagatccaag aatcttcctg tctctgctga tccactgata ggattacaag tacatgccac
     2281 caagcccagc ttcctcttac catctgctgg ggaccaaact taggccctca ttcctacaca
     2341 gtgaatactt gactttgtta tcacccaacc ctaataaata actcactatc caaacaagtt
     2401 gaaaccctta gaattctgtg ttgctccagc atgatgttgt ggtaaacgtt aatacaataa
     2461 gatgcacagg tcataagtgc acattagcta agtgttgaca aagacttaga cctacataac
     2521 ttaaccctat tagccctcca gaaagttcct cattctccat tccaggcaac tttcatcaca
     2581 ccacatcatg tacaactact attgaagttg ttttccacta tagatacaat gagatgtcac
     2641 atacggcttt gtgttttgat ttgcaagtac caatcgagta tgaaatatgg agtggatatt
     2701 ggacattggc caccatctaa atactttgtg ttaaaagaat tggttttcat aatttgtttt
     2761 gtactgactg ctggctagtc agattacctg actagtatgg acaggatttt gcaataatca
     2821 taattctttt ttcggggaac caccacaagg agctcatttt cttgccaaaa gtctggacga
     2881 agccttaaaa cttattgaac aaccagagtt agcagataaa gtggagctgt catggtttgg
     2941 atagttggag gcagttccgt ttacaaggaa gccatgaatc agccaggcca tctcagactc
     3001 tttgtgacaa ggatcatgca ggaatttgaa agtgacacgt tcttcccaga aattgatttg
     3061 gagaaatata aacttctccc agagtaccca ggggtccttt ctgaagtcca ggaggaaaaa
     3121 ggcatcaagt ataaatttga agtctatgag aagaaaggct aacagaaaga tacttgctga
     3181 ttgacttcaa gttctactgc tttcctccta aaattatgca tttttacaag accatgggac
     3241 ttgtgttggc tttagatcta tgagttattc tttctttaga gagggatagt taggaagatg
     3301 tatttgtttt gtggtaccag agatggaacc tgggatcctg tgcatcctgg gcaactgttg
     3361 tactctaagc cactccccaa agtcatgccc cagcccctgt ataattctaa acaattagaa
     3421 ttattttcat tttcattagt ctaaccaggt tatctagagg atccagacat gataagatac
     3481 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3541 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3601 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3661 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctgcaagcct cgtcgtcctg
     3721 gccggaccac gctatctgtg caaggtcccc ggccccggac gcgcgctcca tgagcagagc
     3781 gcccgccgcc gaggcgaaga ctcgggcggc gccctgcccg tcccaccagg tcaacaggcg
     3841 gtaaccggcc tcttcatcgg gaatgcgcgc cttcagcatt gccggcatgt ccccctggcg
     3901 gacgggaagt atccagctcg accaagcttg gcgagatttt caggagctaa ggaagctaaa
     3961 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     4021 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctgcat
     4081 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4141 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4201 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4261 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4321 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4381 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4441 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4501 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4561 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4621 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4681 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4741 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4801 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4861 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4921 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4981 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5041 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5101 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5161 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5221 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5281 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5341 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5401 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5461 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5521 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5581 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5641 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     5701 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5761 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5821 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5881 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5941 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6001 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6061 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6121 ccctttcgtc
//