LOCUS       pE-SUMO_CRYBA4        6323 bp DNA     circular SYN 11-JUN-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6323)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 1, 2024 from SnapGene 7.1.2
            https://www.snapgene.com
COMMENT     Sequence Label: pE-SUMO CRYBA4
FEATURES             Location/Qualifiers
     source          1..6323
                     /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=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..442
                     /label=CRYBA4_NC_F
                     /note="color: black; sequence: GCGGTCTCTAGGTACCCTGCAATG"
     CDS             432..1016
                     /codon_start=1
                     /label=CRYBA4
                     /note="color: #00ff00"
                     /translation="TLQCTKSAGPWKMVVWDEDGFQGRRHEFTAECPSVLELGFETVRS
                     LKVLSGAWVGFEHAGFQGQQYILERGEYPSWDAWGGNTAYPAERLTSFRPAACANHRDS
                     RLTIFEQENFLGKKGELSDDYPSLQAMGWEGNEVGSFHVHSGAWVCSQFPGYRGFQYVL
                     ECDHHSGDYKHFREWGSHAPTFQVQSIRRIQQ"
     primer_bind     complement(1002..1019)
                     /label=CRYBA4_NC_R
                     /note="color: black; sequence: GCTCTAGATTACTGCTGGATCCTGCG"
     CDS             1071..1088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     primer_bind     complement(1098..1122)
                     /label=pE-SUMO_seq_NC_R
                     /note="color: black; sequence: CTTCCTTTCGGGCTTTGTTAGCAGC"
     terminator      1155..1202
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      1239..1694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        1721..1825
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(1826..1894,1895..2686)
                     /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: 1826 .. 1894 / #ccffcc / signal sequence
                        2: 1895 .. 2686 / #ccffcc
                     Cleavage site after base 1894"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2857..3445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3631..3773
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(3875..4066)
                     /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(4875..5957)
                     /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(5958..6035)
                     /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 taccctgcaa tgcacaaagt cagcgggacc ctggaagatg gtggtgtggg
      481 atgaggacgg cttccagggc cggcggcacg agttcacggc cgagtgcccc agcgtgctgg
      541 agcttggctt cgagactgtg cgatctttga aagtgctgag tggagcgtgg gtgggcttcg
      601 agcatgctgg cttccaaggg cagcagtaca ttctggaacg aggcgaatat ccaagctggg
      661 atgcctgggg cggcaacacg gcctaccccg ccgagaggct cacctccttc cggcctgcgg
      721 cctgtgctaa ccaccgtgac tcgaggctga caatcttcga gcaagagaac ttcctgggca
      781 agaaaggaga gctgagcgat gactatcctt ccctccaggc catgggatgg gaaggcaatg
      841 aagtagggtc cttccacgtc cactctgggg cctgggtttg ctcccagttt ccgggctacc
      901 gaggatttca gtatgtgctg gaatgcgatc accattccgg tgactacaaa catttccggg
      961 agtggggctc tcatgccccg accttccagg tgcagagcat ccgcaggatc cagcagtaac
     1021 tagaggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc
     1081 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1141 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1201 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     1261 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1321 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1381 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1441 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1501 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1561 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1621 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1681 aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1741 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1801 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1861 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1921 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1981 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2041 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2101 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2161 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2221 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2281 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2341 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     2401 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2461 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2521 atctggagcc ggtgagcgtg gttctcgcgg tatcattgca gcactggggc cagatggtaa
     2581 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2641 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2701 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2761 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2821 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2881 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2941 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3001 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3061 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3121 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3181 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3241 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3301 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3361 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3421 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3481 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3541 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3601 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3661 gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca
     3721 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3781 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3841 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3901 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3961 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4021 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4081 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4141 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4201 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4261 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4321 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4381 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4441 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4501 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     4561 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4621 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4681 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4741 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4801 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     4861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     4981 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     5041 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     5101 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     5161 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     5221 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     5281 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     5341 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     5401 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     5461 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     5521 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     5581 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     5641 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     5701 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     5761 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     5821 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     5881 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     5941 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     6001 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     6061 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     6121 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     6181 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     6241 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     6301 gatgcgtccg gcgtagagga tcg
//
