LOCUS       sequence-212046-        4384 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 4384)
  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 4384)
  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..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    19..43
                     /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             56..78
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(126..144)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(127..144)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(202..220)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      216..263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(499..520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    639..660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(673..1755)
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1717..1736
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1756..1833)
                     /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     1979..1998
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2154..2173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2273..2295
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2333..2351)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2419..2523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2524..2592,2593..3384)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3555..4143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4044..4063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4297..4314
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt
       61 ttaactttaa gaaggagata tacgtacaga tctgtcgacg cggccgcatg cataagcttg
      121 agtattctat agtgtcacct aaatcccagc ttgatccggc tgctaacaaa gcccgaaagg
      181 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      241 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa cctggcgtaa
      301 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      361 gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atatttgtta aatcagctca
      421 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
      481 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
      541 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacagct tgcatgcctg
      601 caggtcggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
      781 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
      841 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttga
      901 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
      961 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     1021 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     1081 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     1141 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     1201 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     1261 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     1321 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     1381 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     1441 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     1501 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     1561 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     1621 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     1681 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     1741 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     1801 accgcgaaag gttttgcacc attcgatggt gtcgggtacc gagctcgaat ttacgtgaac
     1861 catcacccaa atcaagtttt ttgcggtcga ggtgccgtaa agctctaaat cggaacccta
     1921 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1981 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2041 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcct gatgcggtat
     2101 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     2161 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2221 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2281 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2341 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtca tacactatta
     2761 tcccgtattg acgccgggca agagcaactc ggtcgccggg cgcggtattc tcagaatgac
     2821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3421 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3481 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3661 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4201 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4261 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4321 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4381 tgat
//
