LOCUS       Exported                5945 bp ds-DNA     circular SYN 10-DEC-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  AUTHORS   Arthur de Jong
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 29, 2020 from SnapGene Viewer 5.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5945
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             36..180
                     /label=ITR
                     /note="ITR"
     primer_bind     163..252
                     /label=KI BB FW1 AJ19019
     misc_feature    163..180
                     /label=D sequence
     misc_feature    187..236
                     /label=Multiple cloning site
     promoter        237..1415
                     /label=EF-1-alpha promoter
     primer_bind     312..331
                     /label=oJWseq5 EF1a FW1
     primer_bind     complement(575..591)
                     /label=EF1a rev AJ19025
     primer_bind     1272..1291
                     /label=oJWseq6 EF1a FW2
     primer_bind     complement(1398..1436)
                     /label=BB KI REV1 AJ19020
     primer_bind     1416..1446
                     /label=BB KI FW2 AJ19021
     misc_feature    1416..1425
                     /label=Kozak sequence
     CDS             1428..2132
                     /codon_start=1
                     /label=mCherry
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1433..1445)
                     /label=pEGFP-RP
                     /note="Macrogen Seq primer"
     primer_bind     complement(1840..1866)
                     /label=BB KI REV2 AJ19022
     primer_bind     1847..1872
                     /label=BB KI FW3
     primer_bind     2070..2088
                     /label=oJWseq8 mCherry-C FW
     CDS             2139..2396
                     /codon_start=1
                     /label=KASH TMD
                     /translation="LRSREEEEETDSRMPHLDSPGSSQPRRSFLSRVIRAALPLQLLLL
                     LLLLLACLLPASEDDYSCTQANNFARSFYPMLRYTNGPPPT"
     primer_bind     2444..2462
                     /label=oJWseq9 hGH polyA FW
     primer_bind     complement(2957..2988)
                     /label=BB KI REV3
     misc_feature    2969..2986
                     /label=D sequence
     LTR             2989..3134
                     /label=ITR
                     /label=ITR(1)
                     /note="ITR"
     rep_origin      complement(3378..3966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4137..4997)
                     /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"
     promoter        complement(4998..5102)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5434..5889)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cgctattacg ccaggctgca gggggggggg gggggttggc cactccctct ctgcgcgctc
       61 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
      121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      181 cagatcccat ggggcgcgcc gctagcggta ccggatccta cgtatctaga gtcgacggct
      241 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      301 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      361 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      421 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      481 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      541 ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
      601 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
      661 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
      721 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
      781 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
      841 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
      901 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
      961 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     1021 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
     1081 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     1141 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1201 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     1261 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     1321 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     1381 ttcaaagttt ttttcttcca tttcaggtgt cgtgagccac catgcaagtg agcaagggcg
     1441 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     1501 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1561 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1621 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1681 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     1741 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     1801 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacta
     1861 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     1921 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     1981 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2041 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2101 actccaccgg cggcatggac gagctgtaca agtccggact cagatctcga gaggaggagg
     2161 aggagacaga cagcaggatg ccccacctcg acagccccgg cagctcccag ccgagacgct
     2221 ccttcctctc aagggtgatc agggcagcgc taccgttgca gctgcttctg ctgctgctgc
     2281 tgctcctggc ctgcctgcta cctgcctctg aagatgacta cagctgcacc caggccaaca
     2341 actttgcccg atccttctac cccatgctgc ggtacaccaa cgggccacct cccacctagg
     2401 aattcgatat caagcttatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     2461 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2521 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2581 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2641 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     2701 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     2761 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     2821 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     2881 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     2941 accacgtgcg gaccgagcgg ccgatctgag gaacccctag tgatggagtt ggccactccc
     3001 tctctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     3061 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     3121 actaggggtt ccccctgcag cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3181 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3241 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3301 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3541 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     3601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3781 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3901 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3961 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4021 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4081 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4141 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4201 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4261 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4321 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4381 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4441 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4501 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4561 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4621 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4681 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4741 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4801 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4861 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4921 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4981 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5041 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5101 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5161 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5221 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5281 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     5341 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     5401 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aaacgttaat attttgttaa
     5461 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5521 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5581 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5641 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5701 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5761 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5821 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5881 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5941 ctctt
//
