LOCUS       pE-SUMO_CRYAB        6263 bp DNA     circular SYN 22-JAN-2015
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 29, 2024 from SnapGene 7.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6263
                     /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=CRYAB_NC_F
                     /note="color: black; sequence: GCGAAGACATAGGTATGGACATCG"
     CDS             432..956
                     /codon_start=1
                     /label=CRYAB
                     /note="color: #00ff00"
                     /translation="MDIAIHHPWIRRPFFPFHSPSRLFDQFFGEHLLESDLFPTSTSLS
                     PFYLRPPSFLRAPSWFDTGLSEMRLEKDRFSVNLDVKHFSPEELKVKVLGDVIEVHGKH
                     EERQDEHGFISREFHRKYRIPADVDPLTITSSLSSDGVLTVNGPRKQVSGPERTIPITR
                     EEKPAVTAAPKK"
     primer_bind     complement(943..959)
                     /label=CRYAB_NC_R
                     /note="color: black; sequence: GCTCTAGATTATTTCTTGGGGGCTG"
     misc_feature    957..959
                     /label=Translation stop
                     /note="color: #a6acb3"
     CDS             1011..1028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     primer_bind     complement(1038..1062)
                     /label=pE-SUMO_seq_NC_R
                     /note="color: black; sequence: CTTCCTTTCGGGCTTTGTTAGCAGC"
     terminator      1095..1142
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      1179..1634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        1661..1765
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(1766..1834,1835..2626)
                     /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: 1766 .. 1834 / #ccffcc / signal sequence
                        2: 1835 .. 2626 / #ccffcc
                     Cleavage site after base 1834"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2797..3385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3571..3713
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(3815..4006)
                     /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(4815..5897)
                     /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(5898..5975)
                     /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 tatggacatc gccatccacc acccctggat ccgccgcccc ttctttcctt
      481 tccactcccc cagccgcctc tttgaccagt tcttcggaga gcacctgttg gagtctgatc
      541 ttttcccgac gtctacttcc ctgagtccct tctaccttcg gccaccctcc ttcctgcggg
      601 cacccagctg gtttgacact ggactctcag agatgcgcct ggaaaaggac aggttctctg
      661 tcaacctgga tgtgaagcac ttctccccag aggaactcaa agttaaggtg ttgggagatg
      721 tgattgaggt gcatggaaaa catgaagagc gccaggatga acatggtttc atctccaggg
      781 agttccacag gaaataccgg atcccagctg atgtagaccc tctcaccatt acttcatccc
      841 tgtcatctga tggggtcctc actgtgaatg gaccaaggaa acaggtctct ggccctgagc
      901 gcaccattcc catcacccgt gaagagaagc ctgctgtcac cgcagccccc aagaaataac
      961 tagaggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc
     1021 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1081 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1141 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     1201 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1261 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1321 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1381 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1441 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1501 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1561 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1621 aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1681 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1741 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1801 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1861 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1921 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1981 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2041 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2101 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2161 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2221 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2281 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     2341 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2401 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2461 atctggagcc ggtgagcgtg gttctcgcgg tatcattgca gcactggggc cagatggtaa
     2521 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2581 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2641 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2701 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2941 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3121 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3181 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3241 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3301 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3361 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3421 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3481 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3541 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3601 gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca
     3661 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3721 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3781 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3841 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3901 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     3961 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4021 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4081 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4141 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4201 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4261 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4321 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4381 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4441 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     4501 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4561 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4621 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4681 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4741 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     4801 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4861 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     4921 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4981 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     5041 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     5101 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     5161 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     5221 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     5281 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     5341 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     5401 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     5461 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     5521 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     5581 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     5641 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     5701 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     5761 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     5821 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     5881 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     5941 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     6001 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     6061 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     6121 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     6181 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     6241 gatgcgtccg gcgtagagga tcg
//
