LOCUS       sequence-14339-e        4803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4803)
  AUTHORS   Scheich C, Niesen FH, Seckler R, Bussow K
  TITLE     An automated in vitro protein folding screen applied to a human 
            dynactin subunit.
  JOURNAL   Protein Sci. 2004 Feb . 13(2):370-80.
  PUBMED    14739323
REFERENCE   2  (bases 1 to 4803)
  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: "Protein 
            Sci. 2004 Feb . 13(2):370-80."
COMMENT     Sequence Label: pQTEV
FEATURES             Location/Qualifiers
     source          1..4803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             121..141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             166..186
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      260..354
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             398..1057
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(464..483)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1122..1208
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1301..2383)
                     /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     2345..2364
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2384..2461)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(2545..2567)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2653..2793
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2708..2727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2808..2825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2979..3567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3059..3078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3738..4529,4530..4598))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4361..4380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4599..4703)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4771..4789
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
      121 catcaccatc accatcacca tagcgattac gacatcccca ctactgagaa tctttatttt
      181 cagggatccg ggtcgactgt tgatagatct cggtaccgcg gccgctcgac ctgcagccaa
      241 gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat
      301 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
      361 tagcttggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
      421 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
      481 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
      541 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
      601 tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
      661 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
      721 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
      781 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
      841 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
      901 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
      961 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     1021 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     1081 cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc
     1141 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1201 ggacaaatcc gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg
     1261 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1321 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1381 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     1441 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     1501 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     1561 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     1621 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     1681 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     1741 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     1801 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     1861 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     1921 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     1981 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     2041 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     2101 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     2161 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     2221 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     2281 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     2341 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     2401 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt
     2461 cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc
     2521 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2581 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2641 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     2701 aactatgcgg catcagagca gattgtactg agagtgcacc acatgcggtg tgaaataccg
     2761 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     2821 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2881 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3421 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3481 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3541 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3601 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3661 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3721 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3781 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3841 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3901 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3961 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4021 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4081 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4141 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4201 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4261 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4321 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4381 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4441 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4501 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4561 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4621 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4681 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4741 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     4801 cac
//
