LOCUS       pENTR221-ANXA6-m        5321 bp DNA     circular SYN 26-AUG-2020
DEFINITION  Gateway(R) donor vector with attP1 and attP2 sites and a kanamycin 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  AUTHORS   Thermo Fisher (Invitrogen)
  TITLE     Direct Submission
  JOURNAL   Exported Mar 12, 2024 from SnapGene Viewer 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5321
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..669
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             707..2725
                     /codon_start=1
                     /product="Annexin VI (ANXA6) (NM_001155) Human"
                     /label=Annexin A6
                     /translation="MAKPAQGAKYRGSIHDFPGFDPNQDAEALYTAMKGFGSDKEAILD
                     IITSRSNRQRQEVCQSYKSLYGKDLIADLKYELTGKFERLIVGLMRPPAYCDAKEIKDA
                     ISGIGTDEKCLIEILASRTNEQMHQLVAAYKDAYERDLEADIIGDTSGHFQKMLVVLLQ
                     GTREEDDVVSEDLVQQDVQDLYEAGELKWGTDEAQFIYILGNRSKQHLRLVFDEYLKTT
                     GKPIEASIRGELSGDFEKLMLAVVKCIRSTPEYFAERLFKAMKGLGTRDNTLIRIMVSR
                     SELDMLDIREIFRTKYEKSLYSMIKNDTSGEYKKTLLKLSGGDDDAAGQFFPEAAQVAY
                     QMWELSAVARVELKGTVRPANDFNPDADAKALRKAMKGLGTDEDTIIDIITHRSNVQRQ
                     QIRQTFKSHFGRDLMTDLKSEISGDLARLILGLMMPPAHYDAKQLKKAMEGAGTDEKAL
                     IEILATRTNAEIRAINEAYKEDYHKSLEDALSSDTSGHFRRILISLATGHREEGGENLD
                     QAREDAQVAAEILEIADTPSGDKTSLETRFMTILCTRSYPHLRRVFQEFIKMTNYDVEH
                     TIKKEMSGDVRDAFVAIVQSVKNKPLFFADKLYKSMKGAGTDEKTLTRIMVSRSEIDLL
                     NIRREFIEKYDKSLHQAIEGDTSGDFLKALLALCGGED"
     CDS             2735..3430
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     protein_bind    complement(3446..3545)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3586..3602)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3715..4524
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4671..5259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccccc aaataatgat tttattttga ctgatagtga
      601 cctgttcgtt gcaacaaatt gataagcaat gcttttttat aatgccaact ttgtacaaaa
      661 aagcaggctg gcgccggaac caattcagtc gactggtacc ggcaccatgg ccaaaccagc
      721 acagggtgcc aagtaccggg gctccatcca tgacttccca ggctttgacc ccaaccagga
      781 tgccgaggct ctgtacactg ccatgaaggg ctttggcagt gacaaggagg ccatactgga
      841 cataatcacc tcacggagca acaggcagag gcaggaggtc tgccagagct acaagtccct
      901 ctacggcaag gacctcattg ctgatttaaa gtatgaattg acgggcaagt ttgaacggtt
      961 gattgtgggc ctgatgaggc cacctgccta ttgtgatgcc aaagaaatta aagatgccat
     1021 ctcgggcatt ggcactgatg agaagtgcct cattgagatc ttggcttccc ggaccaatga
     1081 gcagatgcac cagctggtgg cagcatacaa agatgcctac gagcgggacc tggaggctga
     1141 catcatcggc gacacctctg gccacttcca gaagatgctt gtggtcctgc tccagggaac
     1201 cagggaggag gatgacgtag tgagcgagga cctggtacaa caggatgtcc aggacctata
     1261 cgaggcaggg gaactgaaat ggggaacaga tgaagcccag ttcatttaca tcttgggaaa
     1321 tcgcagcaag cagcatcttc ggttggtgtt cgatgagtat ctgaagacca cagggaagcc
     1381 gattgaagcc agcatccgag gggagctgtc tggggacttt gagaagctaa tgctggccgt
     1441 agtgaagtgt atccggagca ccccggaata ttttgctgaa aggctcttca aggctatgaa
     1501 gggcctgggg actcgggaca acaccctgat ccgcatcatg gtctcccgta gtgagttgga
     1561 catgctcgac attcgggaga tcttccggac caagtatgag aagtccctct acagcatgat
     1621 caagaatgac acctctggcg agtacaagaa gactctgctg aagctgtctg ggggagatga
     1681 tgatgctgct ggccagttct tcccggaggc agcgcaggtg gcctatcaga tgtgggaact
     1741 tagtgcagtg gcccgagtag agctgaaggg aactgtgcgc ccagccaatg acttcaaccc
     1801 tgacgcagat gccaaagcgc tgcggaaagc catgaaggga ctcgggactg acgaagacac
     1861 aatcatcgat atcatcacgc accgcagcaa tgtccagcgg cagcagatcc ggcagacctt
     1921 caagtctcac tttggccggg acttaatgac tgacctgaag tctgagatct ctggagacct
     1981 ggcaaggctg attctggggc tcatgatgcc accggcccat tacgatgcca agcagttgaa
     2041 gaaggccatg gagggagccg gcacagatga aaaggctctt attgaaatcc tggccactcg
     2101 gaccaatgct gaaatccggg ccatcaatga ggcctataag gaggactatc acaagtccct
     2161 ggaggatgct ctgagctcag acacatctgg ccacttcagg aggatcctca tttctctggc
     2221 cacggggcat cgtgaggagg gaggagaaaa cctggaccag gcacgggaag atgcccaggt
     2281 ggctgctgag atcttggaaa tagcagacac acctagtgga gacaaaactt ccttggagac
     2341 acgtttcatg acgatcctgt gtacccggag ctatccgcac ctccggagag tcttccagga
     2401 gttcatcaag atgaccaact atgacgtgga gcacaccatc aagaaggaga tgtctgggga
     2461 tgtcagggat gcatttgtgg ccattgttca aagtgtcaag aacaagcctc tcttctttgc
     2521 cgacaaactt tacaaatcca tgaagggtgc tggcacagat gagaagactc tgaccaggat
     2581 catggtatcc cgcagtgaga ttgacctgct caacatccgg agggaattca ttgagaaata
     2641 tgacaagtct ctccaccaag ccattgaggg tgacacctcc ggagacttcc tgaaggcctt
     2701 gctggctctc tgtggtggtg aggacggcgg atccatggtg agcaagggcg aggcagtgat
     2761 caaggagttc atgcggttca aggtgcacat ggagggctcc atgaacggcc acgagttcga
     2821 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2881 gaccaagggt ggccccctgc ccttctcctg ggacatcctg tcccctcagt tcatgtacgg
     2941 ctccagggcc ttcaccaagc accccgccga catccccgac tactataagc agtccttccc
     3001 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac
     3061 ccaggacacc tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa
     3121 cttccctcct gacggccccg taatgcagaa gaagacaatg ggctgggaag cgtccaccga
     3181 gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa
     3241 ggacggcggc cgctacctgg cggacttcaa gaccacctac aaggccaaga agcccgtgca
     3301 gatgcccggc gcctacaacg tcgaccgcaa gttggacatc acctcccaca acgaggacta
     3361 caccgtggtg gaacagtacg aacgctccga gggccgccac tccaccggcg gcatggacga
     3421 gctgtacaag tagtaactcg agtagaccca gctttcttgt acaaagttgg cattataaga
     3481 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     3541 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     3601 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     3661 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     3721 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     3781 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     3841 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     3901 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     3961 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     4021 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     4081 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     4141 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     4201 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     4261 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4321 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     4381 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     4441 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     4501 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     4561 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     4621 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4681 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4741 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4801 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4861 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4921 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4981 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5041 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5101 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5161 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5221 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5281 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
