LOCUS       Exported                6906 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    3XFlag DVL2 (WT)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  AUTHORS   Narimatsu M, Bose R, Pye M, Zhang L, Miller B, Ching P, Sakuma R, 
            Luga V, Roncari L, Attisano L, Wrana JL
  TITLE     Regulation of planar cell polarity by Smurf ubiquitin ligases.
  JOURNAL   Cell. 2009 Apr 17. 137(2):295-307.
  PUBMED    19379695
REFERENCE   2  (bases 1 to 6906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6906
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             241..306
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2546..3168
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2745..2764)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3192..3212)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3197..3526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3377..3512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3439..3458
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3568..3587)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3569..3587)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3601..3617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3601..3617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3614..3636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3625..3641
                     /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(3649..3679)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3832..3849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4003..4591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4083..4102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4762..5622)
                     /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     5385..5404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5623..5727)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5754..6209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5841..5860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6051..6072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6335..6357
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6349..6366
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6350..6366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6527..6906
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattgatc tggtaccatg
      241 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
      301 gacaagcttc ccttcaccat ggcgggtagc agcactgggg gcggtggggt tggggagacg
      361 aaggtgattt accacctgga tgaggaagag actccctacc tggtgaagat ccctgtcccc
      421 gccgagcgca tcaccctcgg cgatttcaag agcgtcctgc agcggcccgc gggcgccaag
      481 tactttttca agtctatgga tcaggatttc ggggtggtga aggaagaaat ttcagatgac
      541 aacgcccgcc tcccctgctt caacggaagg gtggtatcct ggctggtgtc ctcagataat
      601 ccccaacccg agatggcccc tccagtccat gagcctcggg cagaactggc gcctccagcc
      661 ccacctttac ctcctttgcc acccgagagg accagcggca ttggggactc aaggcctcca
      721 tccttccacc ctaatgtgtc cagcagccat gagaatctgg agcctgagac agaaaccgag
      781 tcagtagtgt cactgaggcg ggagcggcct cgcaggagag acagcagtga gcatggcgct
      841 gggggccaca ggactggtgg cccctcaagg ctggagcgcc acctggccgg atacgagagc
      901 tcctctaccc tcatgaccag cgagctggag agtaccagcc tgggggactc ggacgaggag
      961 gacaccatga gcaggttcag cagctccacg gagcagagca gtgcctcccg cctccttaag
     1021 cgccaccggc ggcgaaggaa gcagaggcca ccccgcctgg agaggacgtc atccttcagc
     1081 agcgtcacag attccacaat gtctctcaat atcatcacag tcacgctaaa catggagaag
     1141 tacaacttcc tgggtatctc cattgttggc cagagcaatg agcggggaga cggaggcatc
     1201 tacattggct ccatcatgaa gggtggggct gtggcggccg acgggcgcat tgagccaggg
     1261 gacatgcttt tgcaggtgaa tgacatgaac tttgagaaca tgagcaacga tgacgctgtg
     1321 cgggtgctga gggacattgt gcacaagcct ggccccattg tgctgactgt ggccaagtgc
     1381 tgggatccct ctcctcaggc ctatttcact ctcccccgaa atgagcccat ccagccaatt
     1441 gaccctgctg cctgggtgtc ccattccgcg gctctgactg gcaccttccc agcctatcca
     1501 ggttcctcct ccatgagcac cattacatct ggatcgtctt tgcctgatgg ctgtgaaggc
     1561 cggggtctct ccgtccatac ggacatggca tcggtgacca aggccatggc agctccagag
     1621 tctggactgg aagtccggga ccgcatgtgg ctcaagatca ccatccctaa tgcctttctg
     1681 ggctcggatg tggttgactg gctctaccat cacgtggagg gctttcctga gcggcgggag
     1741 gcccgcaagt atgccagcgg gctgctcaaa gcaggcctga tccgacacac cgtcaacaag
     1801 atcaccttct ctgagcagtg ctattacgtc ttcggagacc tcagtggtgg ctgtgagagc
     1861 tacctagtca acctgtctct caatgacaac gatggctcca gtggggcttc agaccaggat
     1921 accctggctc ctctgcctgg ggccaccccc tggcccctgc tgcccacttt ctcctaccaa
     1981 taccctgccc cacaccccta cagcccgcag cctccaccct accatgagct ttcatcttac
     2041 acctatggtg ggggcagtgc cagcagccag catagtgagg gcagccggag cagtgggtcg
     2101 acacggagtg atgggggggc agggcgcacg gggaggcccg aggagcgggc ccccgagtcc
     2161 aagtccggca gtggcagtga gtctgagccc tccagccgag ggggcagcct tcggcggggt
     2221 ggggaagcaa gtgggactag cgatgggggc cctcctccat ccagaggctc aactgggggt
     2281 gcccctaatc tccgagccca cccagggctc catccctatg gaccgccccc tggcatggcc
     2341 ctcccctaca accccatgat ggtggtcatg atgcccccac ctccacctcc agtccctcca
     2401 gcagtgcagc ctccgggggc ccctccagtc agagacctgg gctctgtgcc cccagaactg
     2461 acagccagcc gccaaagctt ccacatggcc atgggcaatc ccagcgagtt ctttgtggat
     2521 gttatgtgaa ttcaagggcg atcccgggtg gcatccctgt gacccctccc cagtgcctct
     2581 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     2641 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     2701 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     2761 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     2821 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     2881 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     2941 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3001 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     3061 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     3121 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     3181 agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcctcgatcg
     3541 aggaactgaa aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat
     3601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3661 cggaagcata aagtgtaaag cctggggtgc ctaatgagcg agctaactca cattaattgc
     3721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     5761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5821 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5881 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5941 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6061 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6121 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6181 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
     6241 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     6301 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6361 gccagtgcca agctgatcta tacattgaat caatattggc aattagccat attagtcatt
     6421 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     6481 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
     6541 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6601 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6661 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6781 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6841 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6901 accatg
//
