LOCUS       sequence-157317-        5045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the synthetic coiled coil peptide SYNZIP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5045)
  AUTHORS   Thompson KE, Bashor CJ, Lim WA, Keating AE
  TITLE     SYNZIP protein interaction toolbox: in vitro and in vivo 
            specifications of heterospecific coiled-coil interaction domains.
  JOURNAL   ACS Synth Biol. 2012 Apr 20;1(4):118-29.
  PUBMED    22558529
REFERENCE   2  (bases 1 to 5045)
  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: "ACS Synth 
            Biol."; date: "2012-04-20"; volume: "1(4)"; pages: "118-29"
COMMENT     Sequence Label: pQLinkHD-SYNZIP2
FEATURES             Location/Qualifiers
     source          1..5045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(36..50,51..56,57..73,74..80)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    56..72
                     /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    88..104
                     /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             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             147..167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    180..204
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(406..430)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      452..546
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             640..1299
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(706..725)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1364..1450
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1543..2625)
                     /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     2587..2606
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2626..2703)
                     /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(2787..2809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2895..3035
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2950..2969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3050..3067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3221..3809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3301..3320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3980..4771,4772..4840))
                     /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     4603..4622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4841..4945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5013..5031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
       61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
      121 ttaaagagga gaaattaact atgaaacatc accatcacca tcaccatggt gctgcaggta
      181 caagtttgta caaaaaagca ggctccgcgg ccgccccctt caccatggga tccgcgcgta
      241 acgcgtatct gcgtaagaaa atcgcacgtc tgaaaaaaga caacctgcag ctggaacgtg
      301 atgaacagaa cctggaaaaa atcatcgcga acctgcgtga cgaaatcgcg cgtctcgaaa
      361 acgaagttgc gtctcacgaa cagtgataaa agggtgggcg cgccgaccca gctttcttgt
      421 acaaagtggt aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
      481 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
      541 gagaatccaa gtaacaacac catttaaatg gagtggttac aaatggagtg gttaattaag
      601 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
      661 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
      721 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
      781 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
      841 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
      901 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
      961 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1021 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1081 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1141 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1201 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1261 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     1321 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     1381 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1441 taggacaaat ccgccctcta gattacgtgc agtcgatgat aagctgtcaa acatgagaat
     1501 tgtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     1561 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1621 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     1681 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     1741 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     1801 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     1861 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     1921 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     1981 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2041 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2101 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2161 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2221 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     2281 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     2341 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     2401 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     2461 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     2521 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     2581 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     2641 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     2701 gtcggaattt cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcta gagctgcctc
     2761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2881 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     2941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccacatgcgg tgtgaaatac
     3001 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3061 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3121 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3661 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3721 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3781 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3841 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3901 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3961 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4021 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4081 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4141 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4201 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4261 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4321 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4381 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4441 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4501 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4561 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4621 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4681 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4741 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4801 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4861 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4921 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4981 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5041 ttcac
//
