LOCUS       sequence-49948-e        5609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5609)
  AUTHORS   Su PC, Berger BW
  TITLE     Identifying Key Juxtamembrane Interactions in Cell Membranes Using 
            AraTM.
  JOURNAL   J Biol Chem. 2012 Jul 22.
  PUBMED    22822084
REFERENCE   2  (bases 1 to 5609)
  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: "J Biol 
            Chem. 2012 Jul 22."
COMMENT     Sequence Label: pAraTM
FEATURES             Location/Qualifiers
     source          1..5609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             71..1153
                     /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     complement(90..109)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1385..1390,1391..1407,1408..1414)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1422..1438
                     /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)."
     primer_bind     1427..1449
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(1459..1536,1537..2634)
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="MEIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2608..2631
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2635..2661
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3150..3167)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3320..3406
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3498..3525
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3884..4699)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3884..4699)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4607..4626
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4700..4791)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4807..5556)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(4817..4834)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5057..5076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
       61 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      121 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
      181 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
      241 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
      301 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
      361 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
      421 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
      481 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
      541 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
      601 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
      661 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
      721 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
      781 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
      841 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
      901 agacagctca tgttatatcc cgccgtcaac caccatcaaa caggattttc gcctgctggg
      961 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1021 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     1081 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1141 aagcgggcag tgagcgcaac gcaattaatg tgagttagcg cgaattgatc tggtttgaca
     1201 gcttatcatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg
     1261 tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc
     1321 gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga
     1381 gctgttgaca attaatcatc cggctcgtat aatgtgtgga attgtgagcg gataacaatt
     1441 tcacacagga aacagaccat ggaaataaaa acaggtgcac gcatcctcgc attatccgca
     1501 ttaacgacga tgatgttttc cgcctcggct ctcgccaaaa tcgaagaagg taaactggta
     1561 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
     1621 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
     1681 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
     1741 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
     1801 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
     1861 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
     1921 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
     1981 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
     2041 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
     2101 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
     2161 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
     2221 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
     2281 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
     2341 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
     2401 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
     2461 gaagagttgg cgaaagatcc acgtattgcc gccactatgg aaaacgccca gaaaggtgaa
     2521 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     2581 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gacttatccg
     2641 tatgatgtgc cggattatgc ggagctcact agtggatccg aattcggctc cgtcgacaag
     2701 tttgcggccg cactcgagtc tagaggtacc atgaacgagt cgctccatcc accgatggat
     2761 aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat
     2821 atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc
     2881 cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgcattag tcaggcgaag
     2941 ctgcttttga gcactacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac
     3001 gatcaactct atttctcgcg agtatttaaa aaatgcaccg gggccagccc gagcgagttt
     3061 cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata aaagcttggc
     3121 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     3181 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     3241 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     3301 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     3361 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     3421 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     3481 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     3541 tctttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3601 tgataaatgc ttcaataata ttgaaaaaga ctagtggtct tgaggggttt tttgctgaaa
     3661 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     3721 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
     3781 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     3901 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     3961 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     4021 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     4081 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     4141 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     4201 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     4261 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     4321 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     4381 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     4441 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     4501 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     4561 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     4621 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     4681 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     4801 agcgctcttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     4861 cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg gataaagccg
     4921 gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca ggcgtttttc
     4981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca gaggtggcga
     5041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5161 gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cattggtaac
     5341 tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg aaagaacaga
     5401 ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg tagctcagcg
     5461 aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag atgatgaatc
     5521 aatcggtcta tcaagtcaac gaacagctat tccgttactc tagatttcag tgcaatttat
     5581 ctcttcaaat gtagcacctg aagtcacta
//
