LOCUS       sequence-220147-        4408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as non-fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4408)
  AUTHORS   Peranen J, Rikkonen M, Hyvonen M, Kaariainen L
  TITLE     T7 vectors with modified T7lac promoter for expression of proteins 
            in Escherichia coli.
  JOURNAL   Anal Biochem. 1996 May 1;236(2):371-3.
  PUBMED    8660525
REFERENCE   2  (bases 1 to 4408)
  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: "Anal 
            Biochem."; date: "1996-05-1"; volume: "236(2)"; pages: "371-3"
COMMENT     Sequence Label: pBAT4
FEATURES             Location/Qualifiers
     source          1..4408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..111
                     /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."
     CDS             112..1194
                     /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     complement(131..150)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     1344..1365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(1601..1648)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1644..1662
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        1720..1738
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1720..1737
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     RBS             1818..1840
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1853..1877
                     /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)."
     primer_bind     complement(1876..1895)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1877..1895)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1966..1983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2137..2725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2217..2236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2896..3687,3688..3756))
                     /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     3519..3538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3757..3861)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3929..3947
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3985..4007)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4107..4126
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4280..4299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggtgatgg ttcacaattc gagctcggta cccgacacca tcgaatggtg caaaaccttt
       61 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
      121 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
      181 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
      241 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
      301 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
      361 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
      421 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
      481 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
      541 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
      601 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
      661 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
      721 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
      781 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      841 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      901 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
      961 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     1021 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     1081 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     1141 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     1201 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     1261 gacctgcagg catgcaagct gtgggccatc gccctgatag acggtttttc gccctttgac
     1321 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1381 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1441 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1501 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1561 ctattacgcc aggttatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc
     1621 gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag
     1681 cttcctttcg ggctttgtta gcagccggat caagctggga tttaggtgac actatagaat
     1741 actcaagctt atgcatgcgg ccgcatctag agggcccgga tccctcgagg tcgacgaatt
     1801 cgatatccat ggatatatct ccttcttaaa gttaaacaaa attatttcta gaggaattgt
     1861 tatccgctca caattcccta tagtgagtcg tattaatcag ctgcattaat gaatcggcca
     1921 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1981 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2041 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2101 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2161 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2221 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2281 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2341 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2401 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2461 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2521 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2581 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2641 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2701 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2761 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2821 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2881 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2941 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3001 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3061 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3121 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3181 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3241 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3301 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3361 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3421 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3481 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3541 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3601 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3661 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3721 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3781 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3841 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3901 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     3961 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4021 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4081 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4141 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggacgcgccc
     4201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4261 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4321 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4381 cggcacctcg accccaaaaa acttgatt
//
