LOCUS       sequence-366931-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible E coli expression of the Z18 helixCAM..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  AUTHORS   Chao G, Wannier TM, Gutierrez C, Borders NC, Appleton E, Chadha A, 
            Lebar T, Church GM
  TITLE     helixCAM: A platform for programmable cellular assembly in bacteria 
            and human cells.
  JOURNAL   Cell. 2022 Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012.
  PUBMED    36055250
REFERENCE   2  (bases 1 to 5942)
  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: "Cell. 2022 
            Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012."
COMMENT     Sequence Label: pQE80_T5_Z18-ehaA_LacI
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..38
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(62..76,77..82,83..99,100..106)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    82..98
                     /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    114..130
                     /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             149..160
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     sig_peptide     167..232
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     terminator      1451..1545
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1589..2248
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1655..1674)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2313..2399
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2492..3574)
                     /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     3536..3555
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3575..3652)
                     /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(3736..3758)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3844..3984
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3899..3918
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3999..4016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4170..4758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4250..4269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4929..5720,5721..5789))
                     /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     5552..5571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5790..5894)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc acgtcgagaa
       61 atcataaaaa atttatttgc tttgtgagcg gataacaatt ataatagatt caattgtgag
      121 cggataacaa tttcacacag aattcattaa agaggagaaa ttaactatga aatacctgct
      181 gccgaccgct gctgctggtc tgctgctcct cgctgcccag ccggcgatgg ccatgagagg
      241 atcgtcgatc gctgctacgt tagaaaatga cttagcgcgt ctggaaaatg aaaatgctcg
      301 tctggaaaag gacattgcga atctggagcg tgatttggcg aaattggagc gtgaggaggc
      361 ttatttcggt agcggtagcg gctctggatc aggcagcgga tcactcgaga cacctacgcc
      421 gggtccggat ctgaatgtgg ataatgacct gcgaccggag gcgggtagct acattgcgaa
      481 ccttgcagca gcgaatacca tgttcaccac gcgtctgcat gagcgtctgg gtaatacgta
      541 ctataccgac atggtgacgg gtgagcagaa acaaaccact atgtggatgc gccatgaagg
      601 tggtcataat aaatggcgtg atggcagcgg ccagctgaaa acccaaagca atcgctatgt
      661 tctgcaactg ggaggcgatg tcgcgcagtg gagccaaaac ggcagcgacc gctggcatgt
      721 tggggtcatg gcgggatatg gcaacagcga cagcaaaacc atttcctcgc gaaccggtta
      781 tcgtgcaaaa gcgagtgtga acggatatag cacaggcctc tatgccacct ggtatgccga
      841 tgacgagtcg cgtaatggcg cgtatctcga cagttgggcg cagtacagct ggtttgataa
      901 cacagtgaaa ggggatgact tacaaagtga atcctataaa tcaaaaggat ttaccgcttc
      961 actggaagct ggatacaaac acaaattagc tgaatttaat ggcagccagg gaacgcgtaa
     1021 tgaatggtat gttcagccgc aagcacaggt tacctggatg ggagtcaaag ccgataagca
     1081 ccgcgaaagc aacggaaccc tcgttcatag caacggtgat ggcaatgttc aaacccgact
     1141 tggcgtaaaa acctggctga agagccacca taaaatggat gacggtaaat cccgcgagtt
     1201 ccagccgttt gtagaagtga actggctaca taacagtaag gatttcagca ccagtatgga
     1261 tggcgtgtct gtcactcagg atggagcccg aaatattgct gagataaaaa ccggggtgga
     1321 aggacagcta aatgccaacc tgaatgtctg ggggaatgtg ggcgttcagg ttgccgatag
     1381 gggatataat gacacctctg caatggttgg cattaagtgg caattctgaa agcttaatta
     1441 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1501 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc
     1561 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     1621 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     1681 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1741 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     1801 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1861 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1921 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1981 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     2041 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     2101 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     2161 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     2221 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     2281 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
     2341 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2401 cgccctctag attacgtgca gtcgatgata agctgtcaaa catgagaatt gtgcctaatg
     2461 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2581 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     2641 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     2701 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     2761 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     2821 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     2881 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     2941 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     3001 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     3061 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     3121 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     3181 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     3241 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     3301 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     3361 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     3421 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     3481 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     3541 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     3601 ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcggaatttc
     3661 gggcagcgtt gggtcctggc cacgggtgcg catgatctag agctgcctcg cgcgtttcgg
     3721 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3781 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3841 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     3901 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     3961 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4021 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4801 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4861 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4921 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4981 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5041 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5101 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5161 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5221 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5281 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5341 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5401 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5461 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5521 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5581 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5641 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5701 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5761 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5821 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5881 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5941 at
//
