LOCUS       pSUMO-HsSyx3        6309 bp    DNA     circular SYN 17-NOV-2021
DEFINITION  
SOURCE      
  ORGANISM  
COMMENT     SUMMARY:
            E. coli expression clone (T7 promoter) for expression-optimized DNA of human 
            Syx (aa 393-792) with N-terminal His6 + SUMO + TEV protease cleavage site.
            DETAILS:
            -	Protein sequence is MKSS + HHHHHH + GSSMAS + SUMO (aa 
            1-98 of NP_010798.1) + IE + TEV protease cleavage (ENLYFQ*S; * = cleavage site) 
            + N + human Syx (synectin-binding guanine exchange factor; aa 393-792 from 
            accession number NP_001036128.1).
            -	Confers ampicillin-resistance.
            -	Backbone vector is pET His6 Sumo TEV LIC (2S-T) Addgene-29711.
            Construct design by Ryan Boyd and Debra T. Hansen. 
            Cloning by Felicia M. Craciunescu.
FEATURES             Location/Qualifiers
     source          1..6309
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /vntifkey="98"
     promoter        104..208
                     /gene="bla"
                     /vntifkey="30"
                     /label=AmpR\promoter
     CDS             209..1069
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /vntifkey="4"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /vntifkey="33"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     misc_feature    2014..2156
                     /vntifkey="21"
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /vntifkey="4"
                     /label=rop
     promoter        4010..4028
                     /vntifkey="30"
                     /label=T7\promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4117..4139
                     /vntifkey="32"
                     /label=ribosome\binding\site
                     /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4159..4176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /vntifkey="4"
                     /label=6xHis
     CDS             4195..4485
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /vntifkey="4"
                     /label=SUMO\(aa\1-98\of\NP_010798.1)
     CDS             4495..4515
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /vntifkey="4"
                     /label=TEV\protease\cleavage\ENLYFQ*S
     terminator      5859..5906
                     /vntifkey="43"
                     /label=T7\terminator
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
     promoter        complement(6272..6300)
                     /vntifkey="30"
                     /label=tet\promoter
                     /note="E. coli promoter for tetracycline efflux protein gene"
     CDS             4519..5718
                     /vntifkey="4"
                     /label=HsSyx\aa393-792,\optimized
     CDS             4147..5718
                     /vntifkey="4"
                     /label=EXPRESSED\PROTEIN
     primer          4010..4029
                     /vntifkey="27"
                     /label=PRIMER\T7
     primer          complement(5845..5863)
                     /vntifkey="27"
                     /label=PRIMER\T7TERM
     primer          4034..4055
                     /vntifkey="27"
                     /label=PRIMER\37237\seq\F
BASE COUNT     1533 a      1604 c      1782 g      1390 t 
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 
      181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact 
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc 
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc 
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga 
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc 
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg 
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca 
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac 
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg 
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga 
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga 
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc 
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg 
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga 
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg 
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc 
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg 
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt 
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt 
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg 
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat 
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc 
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt 
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat 
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg 
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg 
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc 
     3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc 
     3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc 
     3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc 
     3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc 
     3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc 
     4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa 
     4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat 
     4141 atacatatga aatcttctca ccatcaccat caccatggtt cttctatggc tagcatgtcg 
     4201 gactcagaag tcaatcaaga agctaagcca gaggtcaagc cagaagtcaa gcctgagact 
     4261 cacatcaatt taaaggtgtc cgatggatct tcagagatct tcttcaagat caaaaagacc 
     4321 actcctttaa gaaggctgat ggaagcgttc gctaaaagac agggtaagga aatggactcc 
     4381 ttaagattct tgtacgacgg tattagaatt caagctgatc agacccctga agatttggac 
     4441 atggaggata acgatattat tgaggctcac agagaacaga ttggtgggat cgaggaaaac 
     4501 ctgtacttcc aatccaatca acaagaggcg gtgtgggaac tgctgcacac cgaggcgagc 
     4561 tacatccgta agctgcgtgt gatcattaac ctgttcctgt gctgcctgct gaacctgcag 
     4621 gaaagcggcc tgctgtgcga ggttgaagcg gagcgtctgt ttagcaacat cccggagatt 
     4681 gcgcaactgc atcgtcgtct gtgggcgagc gtgatggcgc cggttctgga aaaagcgcgt 
     4741 cgtacccgtg cgctgctgca gccgggtgac ttcctgaagg gcttcaaaat gtttggtagc 
     4801 ctgtttaaac cgtacatccg ttattgcatg gaggaagagg gctgcatgga gtacatgcgt 
     4861 ggtctgctgc gtgacaacga cctgttccgt gcgtatatta cctgggcgga aaagcacccg 
     4921 cagtgccaac gtctgaaact gagcgatatg ctggcgaagc cgcaccaacg tctgaccaaa 
     4981 tatccgctgc tgctgaaaag cgtgctgcgt aagaccgaag agccgcgtgc gaaagaagcg 
     5041 gtggttgcga tgatcggcag cgtggagcgt ttcattcacc acgttaacgc gtgcatgcgt 
     5101 cagcgtcaag aacgtcagcg tctggcggcg gtggttagcc gtatcgacgc gtacgaagtg 
     5161 gttgagagca gcagcgacga agttgataag ctgctgaaag agtttctgca cctggatctg 
     5221 accgcgccga ttccgggtgc gagcccggaa gaaacccgtc aactgctgct ggaaggcagc 
     5281 ctgcgtatga aagagggtaa agacagcaag atggatgtgt attgcttcct gtttaccgac 
     5341 ctgctgctgg tgaccaaggc ggttaagaaa gcggaacgta cccgtgttat ccgtccgccg 
     5401 ctgctggttg acaaaattgt ttgccgtgaa ctgcgtgatc cgggcagctt cctgctgatc 
     5461 tacctgaacg agtttcacag cgcggtgggt gcgtatacct ttcaggcgag cggtcaagcg 
     5521 ctgtgccgtg gttgggttga caccatttac aacgcgcaga accaactgca acaactgcgt 
     5581 gcgcaggaac cgccgggtag ccagcaaccg ctgcagagcc tggaagagga agaggatgag 
     5641 caagaagagg aagaggaaga ggaagaagag gaagaagaag gcgaagacag cggcaccagc 
     5701 gcggcgagca gcccgaccta ataattggaa gtggataacg gatccgcgat cgcggcgcgc 
     5761 cacctggtgg ccggccggta ccacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa 
     5821 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct 
     5881 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg 
     5941 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg 
     6001 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc 
     6061 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg 
     6121 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg 
     6181 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg 
     6241 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa 
     6301 acatgagaa 
//
