LOCUS       pE-SUMO_CRYGA        6257 bp DNA     circular SYN 11-JUN-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6257)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 1, 2024 from SnapGene 7.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    40..64
                     /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"
     CDS             108..131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=Start/6xHis
                     /note="color: #cc99b2"
                     /translation="MGHHHHHH"
     CDS             132..431
                     /codon_start=1
                     /label=Smt3 fusion protein
                     /note="color: #ff00ff"
                     /translation="GSLQDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTT
                     PLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     primer_bind     351..374
                     /label=pE-SUMO_seq_NC_F
                     /note="color: black; sequence: ATTCAAGCTGATCAGGCCCCTGAA"
     primer_bind     428..441
                     /label=CRYGA_NC_F
                     /note="color: black; sequence: CGGGTCTCTAGGTGGGAAGATCA"
     CDS             432..950
                     /codon_start=1
                     /label=CRYGA
                     /note="color: #00ff00"
                     /translation="GKITFYEDRDFQGRCYNCISDCPNLRVYFSRCNSIRVDSGCWMLY
                     ERPNYQGHQYFLRRGKYPDYQHWMGLSDSVQSCRIIPHTSSHKLRLYERDDYRGLMSEL
                     TDDCACVPELFRLPEIYSLHVLEGCWVLYEMPNYRGRQYLLRPGDYRRYHDWGGADAKV
                     GSLRRVTDLY"
     primer_bind     complement(936..953)
                     /label=CRYGA_NC_R
                     /note="color: black; sequence: 
                     CGGCTCTAGATTAGTACAAATCGGTGAC"
     CDS             1005..1022
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     primer_bind     complement(1032..1056)
                     /label=pE-SUMO_seq_NC_R
                     /note="color: black; sequence: CTTCCTTTCGGGCTTTGTTAGCAGC"
     terminator      1089..1136
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      1173..1628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        1655..1759
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(1760..1828,1829..2620)
                     /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: 1760 .. 1828 / #ccffcc / signal sequence
                        2: 1829 .. 2620 / #ccffcc
                     Cleavage site after base 1828"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2791..3379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3565..3707
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(3809..4000)
                     /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(4809..5891)
                     /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(5892..5969)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggtcatcacc
      121 atcatcatca cgggtccctg caggactcag aagtcaatca agaagctaag ccagaggtca
      181 agccagaagt caagcctgag actcacatca atttaaaggt gtccgatgga tcttcagaga
      241 tcttcttcaa gatcaaaaag accactcctt taagaaggct gatggaagcg ttcgctaaaa
      301 gacagggtaa ggaaatggac tccttaagat tcttgtacga cggtattaga attcaagctg
      361 atcaggcccc tgaagatttg gacatggagg ataacgatat tattgaggct caccgcgaac
      421 agattggagg tgggaagatc accttctacg aggaccgaga ctttcagggt cgctgctaca
      481 attgcatcag tgactgcccc aacctgcggg tctacttcag ccgctgcaac tccatccgag
      541 tagacagcgg ctgctggatg ctctatgagc gtcccaatta ccagggccac cagtacttcc
      601 tgcgccgagg caagtacccc gactatcagc actggatggg cctcagcgac tcggtccaat
      661 cctgccgtat aattcctcat accagctcgc acaagttaag gctgtacgag agagatgact
      721 accgaggcct tatgtctgag ctcactgatg actgcgcctg tgttccagaa ctgttccgtc
      781 tccctgagat ctattccctc cacgtactgg agggctgctg ggtcctctat gaaatgccca
      841 actaccgggg gcggcagtat ctgctgaggc ctggggacta cagaaggtac cacgactggg
      901 ggggtgcaga tgccaaagtc ggctctttga gacgggtcac cgatttgtac taactagagg
      961 atccgaattc gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc
     1021 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     1081 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     1141 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     1201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1381 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1441 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1501 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1561 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     1621 tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1681 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1741 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1801 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1861 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1921 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1981 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2041 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2101 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2161 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2221 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2281 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2341 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2401 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2461 agccggtgag cgtggttctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2521 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2581 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2641 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2701 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2761 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2821 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2881 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2941 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3001 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3061 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3121 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3181 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3241 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3301 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3361 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3421 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3481 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3541 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3601 tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3661 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3721 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3781 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3841 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3901 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3961 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4021 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4081 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4141 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4201 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     4261 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     4321 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     4381 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     4441 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     4501 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4561 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4621 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4681 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4741 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     4801 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4861 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4921 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4981 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     5041 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     5101 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     5161 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     5221 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5281 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5341 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5401 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5461 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5521 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5581 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5641 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5701 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5761 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5821 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5881 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5941 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     6001 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     6061 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     6121 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     6181 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     6241 tccggcgtag aggatcg
//
