LOCUS       Exported                6242 bp DNA     circular SYN 08-OCT-2021
DEFINITION  PH6Sumo-Pyl1 rotated to position 5924.
ACCESSION   .
VERSION     .
KEYWORDS    PH6Sumo-Pyl1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
COMMENT     Vector molecule:  Pyl1/Bam++Sal
               Fragment ends: BamHI and SalI
               Fragment size: 537
            Insert molecule:  Fragment 2
               Fragment ends: SalI and BamHI
               Fragment size: 5705
FEATURES             Location/Qualifiers
     source          1..6242
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..318
                     /label=His6-SUMO tag
                     /note="color: #cc0000; direction: RIGHT"
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             28..318
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="color: #cc99b2
                     Cleavage site after base 318"
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     misc_feature    325..855
                     /label=Pyl1(36-211)
                     /note="color: #cc0000; direction: RIGHT"
     CDS             883..900
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      967..1014
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      1051..1506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        1533..1637
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    1638..2498
                     /label=AmpR
                     /note="color: #cc0000; direction: RIGHT"
     CDS             1638..2498
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 1638 .. 1706 / #ccffcc / signal sequence
                        2: 1707 .. 2498 / #ccffcc
                     Cleavage site after base 1706"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2669..3257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3443..3585
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(3687..3878)
                     /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(4687..5769)
                     /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"
     misc_feature    complement(4687..5646)
                     /label=LacI
                     /note="color: #cc0000; direction: LEFT"
     promoter        complement(5770..5847)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        6156..6174
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    6175..6199
                     /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             6214..6236
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
ORIGIN
        1 atgggtcatc accatcatca tcacgggtcg gactcagaag tcaatcaaga agctaagcca
       61 gaggtcaagc cagaagtcaa gcctgagact cacatcaatt taaaggtgtc cgatggatct
      121 tcagagatct tcttcaagat caaaaagacc actcctttaa gaaggctgat ggaagcgttc
      181 gctaaaagac agggtaagga aatggactcc ttaagattct tgtacgacgg tattagaatt
      241 caagctgatc aggcccctga agatttggac atggaggata acgatattat tgaggctcac
      301 cgcgaacaga ttggaggtgg atccgaattc acccaactct cccaatcaat cgccgagttc
      361 cacacgtacc aactcggtaa cggccgttgc tcatctctcc tagctcagcg aatccacgcg
      421 ccgccggaaa cagtatggtc cgtggtgaga cgtttcgata ggccacagat ttacaaacac
      481 ttcatcaaaa gctgtaacgt gagtgaagat ttcgagatgc gagtgggatg cacgcgcgac
      541 gtgaacgtga taagtggatt accggcgaat acgtctcgag agagattaga tctgttggac
      601 gatgatcgga gagtgactgg gtttagtata accggtggtg aacataggct gaggaattat
      661 aaatcggtta cgacggttca tagatttgag aaagaagaag aagaagaaag gatctggacc
      721 gttgttttgg aatcttatgt tgttgatgta ccggaaggta attcggagga agatacgaga
      781 ttgtttgctg atacggttat tagattgaat cttcagaaac ttgcttcgat cactgaagct
      841 atgaaccgga actaggtcga caagcttgcg gccgcactcg agcaccacca ccaccaccac
      901 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
      961 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1021 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     1081 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1141 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1201 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1261 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1321 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1381 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1441 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     1501 caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1561 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1621 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1681 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1741 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1801 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1861 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1921 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1981 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2041 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2101 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2161 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2221 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2281 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2341 ccggtgagcg tggttctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2401 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2461 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2521 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2581 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2641 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2701 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2761 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2821 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2881 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2941 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3001 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3061 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3121 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3181 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3241 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3301 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3361 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3421 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3481 tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3541 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3601 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3661 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3721 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3781 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3841 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3901 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3961 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4021 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4081 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4141 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4201 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4261 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4321 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
     4381 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4441 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4501 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     4561 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     4621 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     4681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4741 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     4801 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     4861 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     4921 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     4981 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5041 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5101 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5161 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5221 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5281 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5341 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5401 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5461 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     5521 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     5581 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     5641 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     5701 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     5761 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     5821 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     5881 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     5941 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6001 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6061 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6121 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
     6181 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
     6241 cc
//
