LOCUS       sequence-366929-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible E coli expression of the Z17 helixCAM..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  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 5945)
  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_Z17-ehaA_LacI
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..149
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     64..83
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(164..181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(335..923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(415..434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1094..1885,1886..1954))
                     /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     1717..1736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1955..2059)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2127..2145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2169..2183,2184..2189,2190..2206,2207..2213)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2189..2205
                     /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    2221..2237
                     /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             2256..2267
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     sig_peptide     2274..2339
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     terminator      3561..3655
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3699..4358
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3765..3784)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4423..4509
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4602..5684)
                     /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     5646..5665
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5685..5762)
                     /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(5846..5868)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
       61 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
      121 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
      181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      961 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1021 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1081 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1141 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1201 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1261 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1321 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1381 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1441 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1501 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1561 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1621 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1681 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1741 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1801 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1861 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1921 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1981 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2041 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2101 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcacg
     2161 tcgagaaatc ataaaaaatt tatttgcttt gtgagcggat aacaattata atagattcaa
     2221 ttgtgagcgg ataacaattt cacacagaat tcattaaaga ggagaaatta actatgaaat
     2281 acctgctgcc gaccgctgct gctggtctgc tgctcctcgc tgcccagccg gcgatggcca
     2341 tgagaggatc gaacgagaag gaagagttaa aatctaagaa ggcggaattg cgtaatcgta
     2401 tcgagcaatt gaagcagaag cgcgagcagt taaagcaaaa gattgcaaat ttacgcaagg
     2461 aaattgaggc ctacaagggt agcggtagcg gctctggatc aggcagcgga tcactcgaga
     2521 cacctacgcc gggtccggat ctgaatgtgg ataatgacct gcgaccggag gcgggtagct
     2581 acattgcgaa ccttgcagca gcgaatacca tgttcaccac gcgtctgcat gagcgtctgg
     2641 gtaatacgta ctataccgac atggtgacgg gtgagcagaa acaaaccact atgtggatgc
     2701 gccatgaagg tggtcataat aaatggcgtg atggcagcgg ccagctgaaa acccaaagca
     2761 atcgctatgt tctgcaactg ggaggcgatg tcgcgcagtg gagccaaaac ggcagcgacc
     2821 gctggcatgt tggggtcatg gcgggatatg gcaacagcga cagcaaaacc atttcctcgc
     2881 gaaccggtta tcgtgcaaaa gcgagtgtga acggatatag cacaggcctc tatgccacct
     2941 ggtatgccga tgacgagtcg cgtaatggcg cgtatctcga cagttgggcg cagtacagct
     3001 ggtttgataa cacagtgaaa ggggatgact tacaaagtga atcctataaa tcaaaaggat
     3061 ttaccgcttc actggaagct ggatacaaac acaaattagc tgaatttaat ggcagccagg
     3121 gaacgcgtaa tgaatggtat gttcagccgc aagcacaggt tacctggatg ggagtcaaag
     3181 ccgataagca ccgcgaaagc aacggaaccc tcgttcatag caacggtgat ggcaatgttc
     3241 aaacccgact tggcgtaaaa acctggctga agagccacca taaaatggat gacggtaaat
     3301 cccgcgagtt ccagccgttt gtagaagtga actggctaca taacagtaag gatttcagca
     3361 ccagtatgga tggcgtgtct gtcactcagg atggagcccg aaatattgct gagataaaaa
     3421 ccggggtgga aggacagcta aatgccaacc tgaatgtctg ggggaatgtg ggcgttcagg
     3481 ttgccgatag gggatataat gacacctctg caatggttgg cattaagtgg caattctgaa
     3541 agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     3601 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     3661 ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     3721 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     3781 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     3841 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     3901 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     3961 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     4021 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     4081 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     4141 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     4201 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     4261 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     4321 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     4381 gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct
     4441 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     4501 aggacaaatc cgccctctag attacgtgca gtcgatgata agctgtcaaa catgagaatt
     4561 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4621 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4681 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4741 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4801 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     4861 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     4921 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     4981 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     5041 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     5101 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     5161 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5221 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5281 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5341 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5401 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5461 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5521 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5581 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5641 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5701 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg
     5761 tcggaatttc gggcagcgtt gggtcctggc cacgggtgcg catgatctag agctgcctcg
     5821 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5881 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5941 gcggg
//
