LOCUS       pEF.DEST51-ANXA6        8585 bp DNA     circular SYN 26-AUG-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 12, 2024 from SnapGene Viewer 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8585
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        475..1653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          706..1644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1670..1688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1720..1744
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1782..3800
                     /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             3810..4505
                     /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(4521..4545)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4571..4612
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4622..4639
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4668..4892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4938..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5380..5710
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5561..5696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5758..5805
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5824..6222
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6380..6501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6952..7540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7711..8502,8503..8571))
                     /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(join(8572..8585,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgcctc gtgaggctcc
      481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     1021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     1081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1621 caaagttttt ttcttccatt tcaggtgtcg tgaggaatta gcttggtact aatacgactc
     1681 actataggga gacccaagct ggctaggtaa gcttgatcaa caagtttgta caaaaaagca
     1741 ggctggcgcc ggaaccaatt cagtcgactg gtaccggcac catggccaaa ccagcacagg
     1801 gtgccaagta ccggggctcc atccatgact tcccaggctt tgaccccaac caggatgccg
     1861 aggctctgta cactgccatg aagggctttg gcagtgacaa ggaggccata ctggacataa
     1921 tcacctcacg gagcaacagg cagaggcagg aggtctgcca gagctacaag tccctctacg
     1981 gcaaggacct cattgctgat ttaaagtatg aattgacggg caagtttgaa cggttgattg
     2041 tgggcctgat gaggccacct gcctattgtg atgccaaaga aattaaagat gccatctcgg
     2101 gcattggcac tgatgagaag tgcctcattg agatcttggc ttcccggacc aatgagcaga
     2161 tgcaccagct ggtggcagca tacaaagatg cctacgagcg ggacctggag gctgacatca
     2221 tcggcgacac ctctggccac ttccagaaga tgcttgtggt cctgctccag ggaaccaggg
     2281 aggaggatga cgtagtgagc gaggacctgg tacaacagga tgtccaggac ctatacgagg
     2341 caggggaact gaaatgggga acagatgaag cccagttcat ttacatcttg ggaaatcgca
     2401 gcaagcagca tcttcggttg gtgttcgatg agtatctgaa gaccacaggg aagccgattg
     2461 aagccagcat ccgaggggag ctgtctgggg actttgagaa gctaatgctg gccgtagtga
     2521 agtgtatccg gagcaccccg gaatattttg ctgaaaggct cttcaaggct atgaagggcc
     2581 tggggactcg ggacaacacc ctgatccgca tcatggtctc ccgtagtgag ttggacatgc
     2641 tcgacattcg ggagatcttc cggaccaagt atgagaagtc cctctacagc atgatcaaga
     2701 atgacacctc tggcgagtac aagaagactc tgctgaagct gtctggggga gatgatgatg
     2761 ctgctggcca gttcttcccg gaggcagcgc aggtggccta tcagatgtgg gaacttagtg
     2821 cagtggcccg agtagagctg aagggaactg tgcgcccagc caatgacttc aaccctgacg
     2881 cagatgccaa agcgctgcgg aaagccatga agggactcgg gactgacgaa gacacaatca
     2941 tcgatatcat cacgcaccgc agcaatgtcc agcggcagca gatccggcag accttcaagt
     3001 ctcactttgg ccgggactta atgactgacc tgaagtctga gatctctgga gacctggcaa
     3061 ggctgattct ggggctcatg atgccaccgg cccattacga tgccaagcag ttgaagaagg
     3121 ccatggaggg agccggcaca gatgaaaagg ctcttattga aatcctggcc actcggacca
     3181 atgctgaaat ccgggccatc aatgaggcct ataaggagga ctatcacaag tccctggagg
     3241 atgctctgag ctcagacaca tctggccact tcaggaggat cctcatttct ctggccacgg
     3301 ggcatcgtga ggagggagga gaaaacctgg accaggcacg ggaagatgcc caggtggctg
     3361 ctgagatctt ggaaatagca gacacaccta gtggagacaa aacttccttg gagacacgtt
     3421 tcatgacgat cctgtgtacc cggagctatc cgcacctccg gagagtcttc caggagttca
     3481 tcaagatgac caactatgac gtggagcaca ccatcaagaa ggagatgtct ggggatgtca
     3541 gggatgcatt tgtggccatt gttcaaagtg tcaagaacaa gcctctcttc tttgccgaca
     3601 aactttacaa atccatgaag ggtgctggca cagatgagaa gactctgacc aggatcatgg
     3661 tatcccgcag tgagattgac ctgctcaaca tccggaggga attcattgag aaatatgaca
     3721 agtctctcca ccaagccatt gagggtgaca cctccggaga cttcctgaag gccttgctgg
     3781 ctctctgtgg tggtgaggac ggcggatcca tggtgagcaa gggcgaggca gtgatcaagg
     3841 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     3901 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     3961 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     4021 gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     4081 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     4141 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     4201 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     4261 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     4321 gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     4381 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     4441 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     4501 acaagtagta actcgagtag acccagcttt cttgtacaaa gtggttgatc tagagggccc
     4561 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     4621 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     4681 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4741 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4801 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4861 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4921 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5281 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5401 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     5461 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5521 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5641 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5701 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     5761 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     5821 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     5881 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     5941 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     6001 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     6061 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gtcgacaggt gcttctcgat
     6121 ctgcatcctg ggatcaaagc gatagtgaag gacagtgatg gacagccgac ggcagttggg
     6181 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga
     6241 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6301 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6361 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6421 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6481 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7141 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8581 ttttc
//
