LOCUS       sequence-220150-        4376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as non-fusions without 
            initiation ATG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4376)
  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 4376)
  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: pBAT5
FEATURES             Location/Qualifiers
     source          1..4376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        59..77
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    77..101
                     /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             114..136
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(184..202)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(185..202)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(260..278)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      274..321
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(557..578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(728..1810)
                     /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     1772..1791
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1811..1888)
                     /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     2031..2050
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2204..2223)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2323..2345
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2383..2401)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2469..2573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2574..2642,2643..3434)
                     /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     complement(2792..2811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3605..4193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4094..4113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4347..4364
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctgatta
       61 atacgactca ctatagggaa ttgtgagcgg ataacaattc ctctagaaat aattttgttt
      121 aactttaaga aggagatata cgtacagatc tgtcgacgcg gccgcatgca taagcttgag
      181 tattctatag tgtcacctaa atcccagctt gatccggctg ctaacaaagc ccgaaaggaa
      241 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      301 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggataacc tggcgtaata
      361 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
      421 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      481 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      541 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      601 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca cagcttgcat gcctgcaggt
      661 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactta cattaattgc
      721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
      841 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
      901 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
      961 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     1021 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     1081 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     1141 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     1201 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     1261 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     1321 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     1381 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     1441 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     1501 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     1561 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     1621 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     1681 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     1741 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     1801 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     1861 gaaaggtttt gcaccattcg atggtgtcgg gtaccgagct cgaattgtga accatcaccc
     1921 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     1981 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2041 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2101 acacccgccg cgcttaatgc gccgctacag ggcgcgtcct gatgcggtat tttctcctta
     2161 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     2221 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2281 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2341 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2401 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2461 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2521 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2581 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2641 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2701 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2761 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2821 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2881 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2941 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3001 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3061 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3121 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3181 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3241 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3301 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3361 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3421 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3481 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3541 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4201 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4261 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4321 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagag
//
