LOCUS       sequence-14362-e        5288 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 5288)
  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 5288)
  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-PPP1R1A
FEATURES             Location/Qualifiers
     source          1..5288
                     /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      745..839
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             883..1542
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(949..968)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1607..1693
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1786..2868)
                     /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     2830..2849
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2869..2946)
                     /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(3030..3052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3138..3278
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3193..3212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3293..3310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3464..4052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3544..3563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4223..5014,5015..5083))
                     /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     4846..4865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5084..5188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5256..5274
                     /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 agcaagacaa cagcccccga aagatccagt tcacggtccc gctgctggag
      241 ccgcaccttg accccgaggc ggcggagcag attcggaggc gccgccccac ccctgccacc
      301 ctcgtgctga ccagtgacca gtcatcccca gagatagatg aagaccggat ccccaaccca
      361 catctcaagt ccactttggc aatgtctcca cggcaacgga agaagatgac aaggatcaca
      421 cccacaatga aagagctcca gatgatggtt gaacatcacc tggggcaaca gcagcaagga
      481 gaggaacctg agggggccgc tgagagcaca ggaacccagg agtcccgccc acctgggatc
      541 ccagacacag aagtggagtc aaggctgggc acctctggga cagcaaaaaa aactgcagaa
      601 tgcatcccta aaactcacga gagaggcagt aaggaaccca gcacaaaaga accctcaacc
      661 catataccac cactggattc caagggagcc aactcggtct gagcggccgc tcgacctgca
      721 gccaagctta attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac
      781 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
      841 caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
      901 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
      961 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1021 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1081 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1141 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1201 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1261 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     1321 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1381 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     1441 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     1501 gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     1561 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
     1621 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     1681 gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt caaacatgag
     1741 aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     1801 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1861 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1921 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     1981 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     2041 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     2101 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     2161 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     2221 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     2281 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     2341 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     2401 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     2461 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     2521 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2581 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2641 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2701 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     2761 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     2821 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     2881 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc accattcgat
     2941 ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat ctagagctgc
     3001 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3061 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3121 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg cggtgtgaaa
     3241 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     3301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3901 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5221 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5281 gtcttcac
//
