LOCUS       Exported                4190 bp ds-DNA     circular SYN 02-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4190)
  AUTHORS   Weatherall Institute
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 22, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4190
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(448..1308)
                     /codon_start=1
                     /transl_table=11
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /db_xref="GI:672234592"
                     /protein_id="AIJ03088.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1309..1413)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1440..1895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1997..2015
                     
     primer_bind     1999..2020
                     /label=OP_8xCTS_nes_1_FW
     primer_bind     2012..2035
                     /label=OP_8xCTS_nes_2_FW
     misc_feature    2087..2178
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2156..2175
                     /label=P2_test_P3_F
     primer_bind     2186..2200
                     /label=OP_4xCTS_NheI_FW
     misc_feature    2186
                     /label=8x gRNA2 binding sites
                     /note="8x gRNA2 binding sites"
     misc_feature    2186
                     /label=clone
     misc_feature    2187..2192
                     /label=8xCTS_landing_pad
     misc_feature    2201..2220
                     /label=CTS
     misc_feature    2239..2258
                     /label=CTS
     primer_bind     complement(2259..2274)
                     /label=OP_2xCTS_RW
     primer_bind     2270..2284
                     /label=OP_2xCTS_FW
     misc_feature    2285..2304
                     /label=CTS
     misc_feature    2323..2342
                     /label=CTS
     primer_bind     complement(2343..2364)
                     /label=OP_4xCTS_ApaI_RW
     misc_feature    2348..2360
                     /label=8xCTS_landing_pad
     misc_feature    2369..2388
                     /label=CTS
     misc_feature    2407..2426
                     /label=CTS
     primer_bind     complement(2427..2442)
                     /label=OP_2xCTS_RW
     primer_bind     2438..2452
                     /label=OP_2xCTS_FW
     misc_feature    2453..2472
                     /label=CTS
     misc_feature    2491..2510
                     /label=CTS
     primer_bind     complement(2511..2534)
                     /label=OP_4xCTS_ApaI_RW
     misc_feature    2516..2522
                     /label=8xCTS_landing_pad
     misc_feature    2523..2527
                     /label=17nt
     promoter        2528..2559
                     /label=Minimal Ade MLP
                     /note="Minimal Ade MLP"
     primer_bind     complement(2554..2575)
                     /label=P2_clone_P3_R
     primer_bind     complement(2576..2596)
                     /label=OP_8xCTS_nes_2_RW
     CDS             2585..3304
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            2585..3301
                     /gene="ECFP"
                     /label=ECFP
     primer_bind     complement(2603..2623)
                     /label=OP_8xCTS_seq
     primer_bind     complement(3283..3301)
                     
     gene            3321..3542
                     /gene="SV40"
                     /label=SV40
     polyA_signal    3321..3542
                     /gene="SV40"
     polyA_signal    3330..3451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3568..3590)
                     
     rep_origin      complement(join(3879..4190,1..277))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
       61 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      121 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1441 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1501 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1561 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1621 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1681 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1741 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1801 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1861 cgcgaatttt aacaaaatat taacgtttac aatttcccat tcgccattca ggctgcgcaa
     1921 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     1981 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa atatctttat
     2041 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     2101 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     2161 gtgccagaac atttctctat cgatagctag cactagtccc aggacagtac tccgacttac
     2221 ttaatatttt catagccgag gacagtactc cgacttacga gctcctactg accctactag
     2281 tcccaggaca gtactccgac ttacttaata ttttcatagc cgaggacagt actccgactt
     2341 acgagctctc taggggccca ctagtcccag gacagtactc cgacttactt aatattttca
     2401 tagccgagga cagtactccg acttacgagc tcctactgac cctactagtc ccaggacagt
     2461 actccgactt acttaatatt ttcatagccg aggacagtac tccgacttac gagctctcta
     2521 gactagcggg gggctataaa agggggtggg ggcgttcgtc ctgctatcta gcgtcgcgtt
     2581 gaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctggggcgt gcagtgcttc gcccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaacg ccatcagcga caacgtctat atcaccgccg acaagcagaa
     3061 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gtaaggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3361 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3421 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3481 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3541 taaaatcgat aaggatccct cgagtatgtc gaccgatgcc cttgagagcc ttcaacccag
     3601 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     3661 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     3721 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3781 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3841 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3901 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3961 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4021 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     4081 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4141 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
//
