LOCUS       sequence-157322-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the synthetic coiled coil peptide SYNZIP4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  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 5057)
  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-SYNZIP4
FEATURES             Location/Qualifiers
     source          1..5057
                     /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(418..442)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      464..558
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             652..1311
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(718..737)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1376..1462
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1555..2637)
                     /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     2599..2618
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2638..2715)
                     /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(2799..2821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2907..3047
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2962..2981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3062..3079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3233..3821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3313..3332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3992..4783,4784..4852))
                     /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     4615..4634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4853..4957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5025..5043
                     /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 tcccagaaag
      241 ttgcggaact caaaaaccgt gttgcggtta aactgaatcg taacgaacag ctgaaaaaca
      301 aagttgaaga gctgaagaac cgtaacgctt acctcaagaa cgaactggcg accctggaga
      361 acgaggttgc gcgtctggaa aacgacgttg cagaatgata aaagggtggg cgcgccgacc
      421 cagctttctt gtacaaagtg gtaagcttaa ttagctgagc ttggactcct gttgatagat
      481 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
      541 ttttttattg gtgagaatcc aagtaacaac accatttaaa tggagtggtt acaaatggag
      601 tggttaatta aggctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
      661 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
      721 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
      781 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
      841 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
      901 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
      961 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1021 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1081 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1141 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1201 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1261 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     1321 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     1381 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1441 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     1501 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     1561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1621 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     1681 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     1741 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     1801 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     1861 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     1921 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     1981 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2041 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2101 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2161 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2221 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2281 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2341 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2401 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2461 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     2521 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     2581 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     2641 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     2701 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     2761 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2821 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2881 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     2941 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccacatgc
     3001 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3061 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3121 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3181 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3241 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3301 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3361 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3421 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3481 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3541 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3601 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3661 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3721 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3781 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3841 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3901 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3961 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4021 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4081 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4141 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4201 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4261 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4321 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4381 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4441 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4501 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4561 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4621 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4681 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4741 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4801 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4861 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4921 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4981 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5041 ggccctttcg tcttcac
//
