LOCUS       sequence-41839-e        5777 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 5777)
  AUTHORS   Bussow K, Scheich C, Sievert V, Harttig U, Schultz J, Simon B, Bork 
            P, Lehrach H, Heinemann U
  TITLE     Structural genomics of human proteins--target selection and 
            generation of a public catalogue of expression clones.
  JOURNAL   Microb Cell Fact. 2005 Jul 5;4:21. doi: 10.1186/1475-2859-4-21.
  PUBMED    15998469
REFERENCE   2  (bases 1 to 5777)
  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: "Microb Cell
            Fact."; date: "2005-07-5"; volume: "4:21. doi"; pages: " 
            10.1186/1475-2859-4-21"
COMMENT     Sequence Label: pQTEV-MAT2B
FEATURES             Location/Qualifiers
     source          1..5777
                     /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      1234..1328
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1372..2031
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1438..1457)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2096..2182
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2275..3357)
                     /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     3319..3338
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3358..3435)
                     /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(3519..3541)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3627..3767
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3682..3701
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3782..3799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3953..4541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4033..4052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4712..5503,5504..5572))
                     /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     5335..5354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5573..5677)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5745..5763
                     /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 cagggatctg tggggcggga gaaagagctc tctatacact ttgttcccgg gagctgtcgg
      241 ctggtggagg aggaagttaa catccctaat aggagggttc tggttactgg tgccactggg
      301 cttcttggca gagctgtaca caaagaattt cagcagaata attggcatgc agttggctgt
      361 ggtttcagaa gagcaagacc aaaatttgaa caggttaatc tgttggattc taatgcagtt
      421 catcacatca ttcatgattt tcagccccat gttatagtac attgtgcagc agagagaaga
      481 ccagatgttg tagaaaatca gccagatgct gcctctcaac ttaatgtgga tgcttctggg
      541 aatttagcaa aggaagcagc tgctgttgga gcatttctca tctacattag ctcagattat
      601 gtatttgatg gaacaaatcc accttacaga gaggaagaca taccagctcc cctaaatttg
      661 tatggcaaaa caaaattaga tggagaaaag gctgtcctgg agaacaatct aggagctgct
      721 gttttgagga ttcctattct gtatggggaa gttgaaaagc tcgaagaaag tgcagtgact
      781 gttatgtttg ataaagtgca gttcagcaac aagtcagcaa acatggatca ctggcagcag
      841 aggttcccca cacatgtcaa agatgtggcc actgtgtgcc ggcagctagc agagaagaga
      901 atgctggatc catcaattaa gggaaccttt cactggtctg gcaatgaaca gatgactaag
      961 tatgaaatgg catgtgcaat tgcagatgcc ttcaacctcc ccagcagtca cttaagacct
     1021 attactgaca gccctgtcct aggagcacaa cgtccgagaa atgctcagct tgactgctcc
     1081 aaattggaga ccttgggcat tggccaacga acaccatttc gaattggaat caaagaatca
     1141 ctttggcctt tcctcattga caagagatgg agacaaacgg tctttcattg agcggccgct
     1201 cgacctgcag ccaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga
     1261 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     1321 gtgagaatcc aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
     1381 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1441 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1501 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1561 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     1621 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1681 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1741 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1801 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1861 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1921 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1981 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     2041 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     2101 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2161 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     2221 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2341 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2401 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2461 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2521 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2581 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2641 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2701 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2761 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2821 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2881 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2941 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     3001 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     3061 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     3121 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     3181 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3241 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3301 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3361 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     3421 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     3481 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3541 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3601 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3661 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccacatgc
     3721 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3781 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3841 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3901 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3961 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4021 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4081 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4141 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4201 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4261 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4321 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4381 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4441 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4501 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4561 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4621 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4681 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4741 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4801 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4861 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4921 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4981 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5041 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5101 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5161 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5221 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5281 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5341 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5401 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5461 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5521 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5581 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5641 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5701 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5761 ggccctttcg tcttcac
//
