LOCUS       Exported                9042 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEF1a-mRor2WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9042)
  AUTHORS   Mikels A, Minami Y, Nusse R
  TITLE     The Ror2 receptor requires tyrosine kinase activity to mediate Wnt5a
            signaling.
  JOURNAL   J Biol Chem. 2009 Aug 31. ():.
  PUBMED    19720827
REFERENCE   2  (bases 1 to 9042)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9042
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4165..4188
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4207..4236
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4252..4269
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4292..4309)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4298..4522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4568..4996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4655..4674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4865..4886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5005..5025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5010..5339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5190..5325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5252..5271
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5406..6200
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5460..5479)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6070..6089
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6376..6497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6413..6432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6467..6486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6546..6562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6546..6562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6559..6581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6570..6586
                     /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(6594..6624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6639..6660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6777..6794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6937..7525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7017..7036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7696..8556)
                     /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     8319..8338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8557..8661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8736..8755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     1321 accatggctc ggggctgggt gcggccgagc cgtgtgcctc tgtgcgcccg ggccgtctgg
     1381 acggctgcgg cgctcctgct ctggacaccc tggacggcag gtgaagtgga agattcggag
     1441 gcaatcgaca ccttgggaca acctgatgga ccggacagcc cacttcccac tctgaaaggc
     1501 tactttctga attttctgga gccagtcaac aatatcacca ttgttcaggg ccagacggca
     1561 atcctgcact gcaaggtggc gggaaaccca cctcccaatg tgcggtggct gaagaatgat
     1621 gccccggttg tgcaagagcc acgaagggtc gtcatccgga agacagaata cggctcccgg
     1681 ctgcggatcc aagacctgga cacaacagac acaggctact accagtgtgt ggctaccaac
     1741 gggctgaaga ccatcactgc cactggggtt ctatatgtgc ggctcggtcc gacgcacagc
     1801 ccgaaccaca attttcagga tgacgatcag gaagatggct tctgccagcc gtaccgaggg
     1861 atcgcttgtg cgcgcttcat tgggaaccgg actatttatg tggactccct ccagatgcag
     1921 ggggagattg aaaaccgaat cacagctgcc ttcaccatga tcggcacctc cacgcaactg
     1981 tcagaccagt gttcacagtt tgccatccca tccttctgcc acttcgtctt ccctctgtgc
     2041 gacgcatgct cccgggcgcc caagcctcgc gaactgtgcc gggatgaatg tgaggtgctg
     2101 gagaacgacc tgtgccgcca ggagtacacc atcgcccgct ccaacccgct catcctcatg
     2161 cggctccagc tgcccaagtg cgaagcgctg cccatgcccg agagcccgga tgctgcgaac
     2221 tgcatgcgca tcgggatccc cgcggagagg ctgggtcgct accaccagtg ctacaacggc
     2281 tccggcgccg attacagggg gatggccagt accaccaagt caggccacca gtgtcagcct
     2341 tgggctctgc agcaccccca cagccatcgc ctatccagca cggaattccc tgagctggga
     2401 ggaggccatg cctactgccg gaaccccggg ggccagatgg aaggcccgtg gtgctttacg
     2461 cagaataaaa acgtacgcgt ggaactgtgt gacgtacccc cgtgtagtcc ccgagatggc
     2521 agcaagatgg ggattctgta catcctggtc cccagcattg ctatccccct ggtcatcgct
     2581 tgcctgttct tcctcgtctg catgtgccgc aacaaacaga aggcttcggc ctccacccca
     2641 cagcgccggc agctgatggc ctctcccagc caggacatgg agatgccact catcagccag
     2701 cacaaacagg ccaaactcaa agagatcagc ttgtccacag tgaggttcat ggaggagctc
     2761 ggggaggacc ggtttggcaa ggtctacaaa ggccacctgt tcgggcctgc cccaggagaa
     2821 ccaacccagg ccgtggccat caagacgctg aaagacaagg ctgaggggcc cctgcgggag
     2881 gagttccggc aagaggcgat gctccgggcc cgactgcagc accccaacat cgtctgtctc
     2941 ctaggcgtcg tgaccaagga ccaacccttg agcatgatct tcagctactg ttcccatggc
     3001 gaccttcatg aattcctggt catgcgctcg ccgcactccg atgtgggcag caccgatgac
     3061 gaccgcacag tgaagtcagc cctggagccc ccggacttcg tgcacgtggt ggcgcagatc
     3121 gctgcgggga tggagttcct gtccagccac cacgtggtcc ataaggacct ggccacacgc
     3181 aatgtgctgg tgtacgacaa gctgaacgtg aggatctcag acttgggcct cttccgtgag
     3241 gtatactccg cagattacta caaactcatg ggcaattcac tgctgcccat ccgctggatg
     3301 tcccccgagg ccgtcatgta tggaaagttc tccatcgact ctgacatctg gtcctacggt
     3361 gtggtcctct gggaggtctt tagctacggc ctgcagccct actgtgggta ctccaaccag
     3421 gacgtggtgg agatgatccg gagccggcag gtgctgccct gcccggatga ctgccccgcc
     3481 tgggtctatg ccctcatgat tgaatgctgg aatgagttcc caagccggag gccccgcttt
     3541 aaggacatcc acagccggct ccggtcctgg ggcaacctat ccaactataa tagttccgcg
     3601 cagacctcag gagccagcaa caccacacag accagctccc tgagcaccag ccccgtaagc
     3661 aatgtgagca atgcccgcta tatggccccc aagcagaagg cccagccctt cccacagcct
     3721 cagttcatcc ccatgaaggg tcagatcaga cccttggtgc cccccgcaca gctgtacatc
     3781 ccggtgaacg gctatcagcc ggtaccggca tacggggcct acctgcccaa cttctaccca
     3841 gtccagatcc ccatgcagat ggccccacag caggtgcccc ctcagatggt ccccaagccg
     3901 agctcacacc acagtggcag cggctccacc agcactggct acgtcaccac ggcgccctcc
     3961 aatacatctg tggcggacag ggcggcccta ctctctgagg gcaccgagga tgtacagaac
     4021 atcgcggaag acgtggccca gagccctgtg caggaagcag aggaggagga ggaggggtct
     4081 gtccctgaga ctgaactcct gggagacaat gacacgctcc aggtgaccga ggcggctcat
     4141 gtccagcttg aagccctcga gggggactat aaggacgatg atgacaaata atctagaggg
     4201 cccttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg tcatcatcac
     4261 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4321 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4381 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4441 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4501 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4561 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4621 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4681 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4741 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4801 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4861 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4921 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4981 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5041 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5101 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5161 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5221 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5281 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5341 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5401 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5461 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5521 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5581 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5641 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5701 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5761 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5821 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5881 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5941 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6001 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6061 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6121 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6181 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6241 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6301 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6361 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6421 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6481 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6541 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6601 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8701 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     8761 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     8821 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     8881 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
     8941 actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt
     9001 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa tt
//
