LOCUS       sequence-405315-        5921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant protein production .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5921)
  AUTHORS   Posner M, Kiss AJ, Skiba J, Drossman A, Dolinska MB, Hejtmancik JF, 
            Sergeev YV
  TITLE     Functional validation of hydrophobic adaptation to physiological 
            temperature in the small heat shock protein alphaA-crystallin.
  JOURNAL   PLoS One. 2012;7(3):e34438. doi: 10.1371/journal.pone.0034438. Epub 
            2012 Mar 29.
  PUBMED    22479631
REFERENCE   2  (bases 1 to 5921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2012"; volume: "7(3):e34438. doi"; pages: " 
            10.1371/journal.pone.0034438. Epub 2012 Mar 29"
COMMENT     Sequence Label: Danio rerio cryabb
FEATURES             Location/Qualifiers
     source          1..5921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     113..132
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        113..131
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    132..156
                     /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)."
     RBS             171..193
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             765..782
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(835..853)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      849..896
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      933..1388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1020..1039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1230..1251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1414..1518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1519..1587,1588..2379)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1737..1756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3039..3058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3292..3309
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3324..3463
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3389..3408)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3549..3571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3565..3756)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4565..5647)
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5609..5628
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5648..5725)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
       61 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
      121 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      181 tttaagaagg agatatacat atggacatcg ccatcaatcc cccgttccgg cgcatcttat
      241 ttcctatctt ctttccccgc cggcaatttg gggagcatat tactgaggcc gacgtgatct
      301 cctcattgta ctcccaacga tcttcctttc ttcgcagtcc aagttggatg gaaagcggag
      361 tttctgaggt gaaaatggaa aaggaccaat tctcactaag tctggatgtt aaacactttg
      421 caccagagga gctatcagtg aaaatcattg gagatttcat tgaaattcat gctaaacatg
      481 aagatcgtca ggacggtcac ggttttgtct cacgagagtt tttgagaaaa taccgtgtcc
      541 ctgttggtgt ggatccagcc tccatcacgt cctcgctgtc ttctgatggt gtgctgacag
      601 tgacaggccc actaaagctc tcagacggcc cagaacgcac catcgccatc cccgtaactc
      661 gtgatgacaa aaccaccgtc gctggccctc agaagtaaag atgctaaacg tcaactttca
      721 aaaaaaaaaa aaaaaagggc ggccgctcta gagtatccct cgagcaccac caccaccacc
      781 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      841 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      901 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
      961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1021 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1081 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1141 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1201 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1261 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1321 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     1381 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1441 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1501 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1621 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1681 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1741 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1801 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1921 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2041 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     2101 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2341 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2401 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2761 gctctgswrm tsstkwyass akkkgctgct gccagtggcg ataagtcgtg tcttaccggg
     2821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3241 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3301 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3361 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3421 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3481 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3541 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3601 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3661 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3721 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     3781 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3841 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3901 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3961 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     4021 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     4081 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     4141 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     4201 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     4261 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4321 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4381 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4441 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     4501 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     4561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     4681 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     4741 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     4801 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     4861 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     4921 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     4981 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     5041 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     5101 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     5161 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     5221 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     5281 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     5341 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     5401 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     5461 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     5521 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     5581 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     5641 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     5701 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     5761 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     5821 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     5881 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat c
//
