LOCUS       Exported                5159 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCS2 Flag-JunB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5159)
  AUTHORS   Taelman VF, Dobrowolski R, Plouhinec JL, Fuentealba LC, Vorwald PP, 
            Gumper I, Sabatini DD, De Robertis EM
  TITLE     Wnt signaling requires sequestration of glycogen synthase kinase 3 
            inside multivesicular endosomes.
  JOURNAL   Cell. 2010 Dec 23. 143(7):1136-48.
  PUBMED    21183076
REFERENCE   2  (bases 1 to 5159)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5159
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1078..1087
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1087..1110
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2167..2184)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2189..2323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2213..2232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2267..2286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2436..2456)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2436..2454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2475..2491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2475..2491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2488..2510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2499..2515
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2523..2553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2568..2589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2706..2723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2877..3465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2957..2976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3636..4496)
                     /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     4259..4278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4497..4601)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4627..5082)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4764..4785)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4976..4995
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcgcc
     1081 accatggact acaaggacga cgatgacaag tgcactaaaa tggaacagcc cttctaccac
     1141 gacgactcat acacagctac gggatacggc cgggcccctg gtggcctctc tctacacgac
     1201 tacaaactcc tgaaaccgag cctggcggtc aacctggccg acccctaccg gagtctcaaa
     1261 gcgcctgggg ctcgcggacc cggcccagag ggcggcggtg gcggcagcta cttttctggt
     1321 cagggctcgg acaccggcgc gtctctcaag ctcgcctctt cggagctgga acgcctgatt
     1381 gtccccaaca gcaacggcgt gatcacgacg acgcctacac ccccgggaca gtacttttac
     1441 ccccgcgggg gtggcagcgg tggaggtgca gggggcgcag ggggcggcgt caccgaggag
     1501 caggagggct tcgccgacgg ctttgtcaaa gccctggacg atctgcacaa gatgaaccac
     1561 gtgacacccc ccaacgtgtc cctgggcgct accggggggc ccccggctgg gcccgggggc
     1621 gtctacgccg gcccggagcc acctcccgtt tacaccaacc tcagcagcta ctccccagcc
     1681 tctgcgtcct cgggaggcgc cggggctgcc gtcgggaccg ggagctcgta cccgacgacc
     1741 accatcagct acctcccaca cgcgccgccc ttcgccggtg gccacccggc gcagctgggc
     1801 ttgggccgcg gcgcctccac cttcaaggag gaaccgcaga ccgtgccgga ggcgcgcagc
     1861 cgggacgcca cgccgccggt gtcccccatc aacatggaag accaagagcg catcaaagtg
     1921 gagcgcaagc ggctgcggaa ccggctggcg gccaccaagt gccggaagcg gaagctggag
     1981 cgcatcgcgc gcctggagga caaggtgaag acgctcaagg ccgagaacgc ggggctgtcg
     2041 agtaccgccg gcctcctccg ggagcaggtg gcccagctca aacagaaggt catgacccac
     2101 gtcagcaacg gctgtcagct gctgcttggg gtcaagggac acgccttctg actcgagcct
     2161 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     2221 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2281 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2341 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     2401 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2521 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3301 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4621 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4681 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4741 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4801 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4861 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4921 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4981 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5041 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     5101 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacc
//
