LOCUS       pET_SUMO_FL_rSyt        6908 bp DNA     circular SYN 06-DEC-2023
DEFINITION  Chapman_J8Q_10_SUMO_FL_Syt1     coverage: 762x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6908)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 6, 2023 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6908
                     /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..5156
                     /codon_start=1
                     /label=rSyt1
                     /note="color: #993366"
                     /translation="MVSASHPEALAAPVTTVATLVPHNATEPASPGEGKEDAFSKLKQK
                     FMNELHKIPLPPWALIAIAIVAVLLVVTCCFCVCKKCLFKKKNKKKGKEKGGKNAINMK
                     DVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEEKLGKLQYSLDYDFQNNQLLVGII
                     QAAELPALDMGGTSDPYVKVFLLPDKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGK
                     TLVMAVYDFDRFSKHDIIGEFKVPMNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLR
                     YVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYN
                     ESFSFEVPFEQIQKVQVVVTVLDYDKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRR
                     PIAQWHTLQVEEEVDAMLAVKK"
     misc_feature    4311..4670
                     /label=C2A
                     /note="color: #0000ff"
     misc_feature    4704..5105
                     /label=C2B
                     /note="color: #0000ff"
     CDS             5215..5232
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      5299..5346
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      5383..5838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(5930..6745)
                     /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(5930..6745)
                     /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(6867..6908,1..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 atggtgagtg
     3901 ccagtcatcc tgaggccctg gccgcccctg tcaccactgt tgcgaccctt gtcccacaca
     3961 atgccactga gccagccagt cctggggaag ggaaggaaga tgccttttcc aagctgaagc
     4021 agaagtttat gaatgagctg cataaaattc cattgccacc gtgggcctta atagccatag
     4081 ccatagttgc ggtcctttta gtcgtaacct gctgcttttg tgtctgtaag aaatgtttgt
     4141 tcaaaaagaa aaacaagaag aaggggaagg aaaagggagg aaagaacgcc attaacatga
     4201 aagacgtgaa agacttaggg aagaccatga aggatcaggc ccttaaggat gacgatgctg
     4261 aaaccggact gactgatgga gaagaaaagg aagagcccaa ggaagaggag aaactgggaa
     4321 agctccaata ttcactggac tatgacttcc agaataacca gctgttggtg ggaatcatcc
     4381 aggctgctga actgcccgcc ctggacatgg ggggtacatc cgatccatac gtcaaagtct
     4441 tcctgctgcc tgacaaaaag aagaaatttg agactaaagt ccaccggaaa accctcaatc
     4501 cagtcttcaa tgaacaattt actttcaagg taccctactc ggaattaggt ggcaaaaccc
     4561 tggtgatggc tgtgtatgac tttgatcgct tctccaagca cgacatcatc ggagagttca
     4621 aagttcctat gaacaccgtg gattttggcc atgtgaccga ggagtggcgc gatctccaga
     4681 gcgctgagaa agaagagcaa gagaaactgg gtgacatctg cttctccctc cgctacgtcc
     4741 ctactgccgg caaactgact gttgtcattc tggaagccaa gaacctgaag aagatggatg
     4801 tgggtggctt atctgatccc tacgtgaaga ttcacctgat gcagaacggt aagaggctga
     4861 agaagaaaaa gacgacgatt aagaagaaca cactcaaccc ctactacaac gagtccttca
     4921 gctttgaagt tccgttcgag caaatccaga aagtgcaagt ggtggtaact gttttggact
     4981 atgacaagat tggcaagaac gacgccatcg gaaaagtctt cgttggttac aacagcactg
     5041 gggcggagct gcgacactgg tcagacatgc tggccaaccc ccggcgaccc atcgcacagt
     5101 ggcacactct gcaggtagag gaggaggttg atgccatgct ggctgtcaag aagtaaaagc
     5161 ttaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttagt cgagcaccac
     5221 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     5281 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     5341 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     5401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5821 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5881 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     5941 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     6001 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     6061 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     6121 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     6181 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     6241 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     6301 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     6361 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     6421 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     6481 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     6541 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     6601 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     6661 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     6721 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     6781 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6901 ttgcaaac
//
