LOCUS       pET_SUMO_Syt1_S7        6623 bp DNA     circular SYN 06-DEC-2023
DEFINITION  Chapman_J8Q_6_SUMO_Syt1_s7+     coverage: 120x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 6, 2023 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    733..872
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(974..1165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1974..3056)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3057..3134)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        3443..3461
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    3462..3486
                     /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)."
                     /note="color: #31849b"
     RBS             3501..3523
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             3543..3560
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             3570..3587
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /note="color: #cc99b2
                     Cleavage site after base 3581"
                     /translation="LVPRGS"
     CDS             3594..3887
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="color: #cc99b2
                     Cleavage site after base 3887"
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             3891..4871
                     /codon_start=1
                     /label=rSyt1
                     /note="color: #993366"
                     /translation="GGKNAINMKDVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEE
                     KLGKLQYSLDYDFQNNQLLVGIIQAAELPAKDMGGTSDPYVKVFLLPDKKKKFETKVKR
                     KTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSRHDIIGEFKVPMNTVDFGHVTEE
                     WRDLQSAEKEEQEKLGDIAFSLRYVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLM
                     QNGKRLKKKKTTIKKRTLNPYYNESFSFEVPFEQIQKVQVVVTVLDKDKCGRNDAIGKV
                     FVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK"
     misc_feature    4026..4385
                     /label=C2A
                     /note="color: #0000ff"
     misc_feature    4116..4118
                     /label=L171K
                     /note="color: #ff0000"
     misc_feature    4197..4199
                     /label=H198K
                     /note="color: #ff0000"
     misc_feature    4311..4313
                     /label=K236R
                     /note="color: #ff0000"
     misc_feature    4419..4820
                     /label=C2B
                     /note="color: #0000ff"
     misc_feature    4434..4436
                     /label=C277A
                     /note="color: #ff0000"
     misc_feature    4602..4604
                     /label=N333R
                     /note="color: #ff0000"
     misc_feature    4695..4697
                     /label=Y364K
                     /note="color: #ff0000"
     misc_feature    4704..4706
                     /label=I367C
                     /note="color: #ff0000"
     misc_feature    4710..4712
                     /label=K369R
                     /note="color: #ff0000"
     CDS             4930..4947
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      5014..5061
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      5098..5553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(5645..6460)
                     /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"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5645..6460)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      join(6582..6623,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      781 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      841 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      901 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      961 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     1021 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     1081 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     1141 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     1201 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     1261 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     1321 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     1381 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     1441 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     1501 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     1561 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1621 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     1681 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     1741 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     1801 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     1861 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     1921 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     1981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2041 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     2101 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2161 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     2221 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     2281 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     2341 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     2401 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     2461 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     2521 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     2581 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     2641 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     2701 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     2761 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     2821 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     2881 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     2941 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3001 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3061 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     3121 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     3181 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     3241 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     3301 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     3361 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     3421 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     3481 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat atgggcagca
     3541 gccatcatca tcatcatcac ggcagcggcc tggtgccgcg cggcagcgct agcatgtcgg
     3601 actcagaagt caatcaagaa gctaagccag aggtcaagcc agaagtcaag cctgagactc
     3661 acatcaattt aaaggtgtcc gatggatctt cagagatctt cttcaagatc aaaaagacca
     3721 ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca gggtaaggaa atggactcct
     3781 taagattctt gtacgacggt attagaattc aagctgatca gacccctgaa gatttggaca
     3841 tggaggataa cgatattatt gaggctcaca gagaacagat tggtggatcc ggaggaaaga
     3901 acgccattaa catgaaagac gtgaaagact tagggaagac catgaaggat caggccctta
     3961 aggatgacga tgctgaaacc ggactgactg atggagaaga aaaggaagag cccaaggaag
     4021 aggagaaact gggaaagctc caatattcac tggactatga cttccagaat aaccagctgt
     4081 tggtgggaat catccaggct gctgaactgc ccgccaagga catggggggt acatccgatc
     4141 catacgtcaa agtcttcctg ctgcctgaca aaaagaagaa atttgagact aaagtcaagc
     4201 ggaaaaccct caatccagtc ttcaatgaac aatttacttt caaggtgccc tactcggaat
     4261 taggtggcaa aaccctggtg atggctgtgt atgactttga tcgcttctcc aggcatgaca
     4321 tcatcggaga gttcaaagtt cctatgaaca ccgtggattt tggccatgtg accgaggagt
     4381 ggcgtgatct ccagagcgct gagaaagaag agcaagagaa actgggtgac atcgccttct
     4441 ccctccgcta cgtccctact gccggcaaac tgactgttgt cattctggaa gccaagaacc
     4501 tgaagaagat ggatgtgggt ggcttatctg atccctacgt gaagattcac ctgatgcaga
     4561 acggtaagag gctgaagaag aaaaagacga cgattaagaa gaggacactc aacccctact
     4621 acaacgagtc cttcagcttt gaagttccgt tcgagcaaat ccagaaagtg caagtggtgg
     4681 taactgtttt ggacaaggac aagtgtggca ggaacgacgc catcggcaaa gtcttcgttg
     4741 gttacaacag cactggggcg gagctgcgac actggtcaga catgctggcc aacccccggc
     4801 gacccatcgc acagtggcac actctgcagg tagaggagga ggttgatgcc atgctggctg
     4861 tcaagaagta aaagcttagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac
     4921 ttagtcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     4981 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     5041 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     5101 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5161 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5221 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5281 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5341 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5401 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5461 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5521 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     5581 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     5641 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     5701 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     5761 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     5821 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     5881 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     5941 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     6001 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     6061 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     6121 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     6181 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     6241 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     6301 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     6361 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     6421 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     6481 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     6541 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6601 cgcgtaatct gctgcttgca aac
//
