LOCUS       sequence-449992-        5772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone plasmid for genomic insertions into H. neapolitanus 
            neutral site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5772)
  AUTHORS   Turnsek JB, Oltrogge LM, Savage DF
  TITLE     Conserved and repetitive motifs in an intrinsically disordered 
            protein drive ⍺-carboxysome assembly.
  JOURNAL   J Biol Chem. 2024 Jul 4;300(8):107532. doi: 
            10.1016/j.jbc.2024.107532.
  PUBMED    38971311
REFERENCE   2  (bases 1 to 5772)
  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."; date: "2024-07-4"; volume: "300(8):107532. doi"; pages: " 
            10.1016/j.jbc.2024.107532"
COMMENT     Sequence Label: pSAV108
FEATURES             Location/Qualifiers
     source          1..5772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      198..786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     687..706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1614..1633)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1625..2440
                     /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             1625..2440
                     /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     complement(1698..1717)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2584..2606
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2684..2761
                     /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."
     CDS             2762..3844
                     /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     complement(2781..2800)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4076..4081,4082..4098,4099..4105)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4113..4129
                     /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    4224..4245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4260..4265,4266..4283,4284..4290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4298..4314
                     /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     4322..4338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4322..4338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4334..4657
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    4351..4407
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4408..4425)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4408..4424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4417..4439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4633..4652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4772..4789)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4942..5028
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5120..5147
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctcagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
       61 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
      121 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
      181 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      241 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      301 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
      361 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      421 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      481 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      541 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      601 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      661 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      721 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      781 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      841 cacatgttct ttcctgcgtt aggacccaac gctgcccgag atgcgccgcg tgcggctgct
      901 ggagatggcg gacgcgatgg atatgttctg ccacctgcga ctcaaacttc tgatctggga
      961 tggcacgggg gtgtggctct gtcagcgccg gttgcaccgg gggcattttt attggccaaa
     1021 gcccggggat gatcggttta tcatgagtgc cgaacactgg cagtggctga ccaccggggt
     1081 tgactggcag aggctctgcg ccaagccaca ggcccactgg caagtgtgag atttttcagg
     1141 atttcgtctt gaaaaatcag cacatgcagc ctggggtttg gtataatcaa cctcatgatt
     1201 tcaaacgcag aaaccggtca ttcatcccct gaaacgccct ccgtggcgtg gctgattgcg
     1261 caactggaac tgcgggattc tgagttggca aaacttgatg cggcactggt gcaacgcaat
     1321 caagacatcc gccatttaac ccaaaatctt catgccaaag acatcaagat caaggcactc
     1381 acccatgaac tggcgtatta ccgccgtatc cgcttcgggg ccaaaaccga ggccatgagt
     1441 gccgagcaac tgcacctgtt cgaagacgat ctggccgagg tagtaagctt agatcgacct
     1501 gcaggggggg ggggaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1561 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1621 tgttatgagc catattcaac gggaaacgtc ttgctcgagg ccgcgattaa attccaacat
     1681 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1741 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1801 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1861 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1921 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1981 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2041 tccttttaac agcgatcgcg tatttcgact cgctcaggcg caatcacgaa tgaataacgg
     2101 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2161 gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt
     2221 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2281 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2341 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2401 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2461 gtaacactgg cagagcatta cgctgacttg acgggacggc ggctttgttg aataaatcga
     2521 acttttgctg agttgaagga tcagatcacg catcttcccc ttacagacaa gctgtgaccg
     2581 cctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2641 agatcaattc gcgcgcgaag gcgaagcggc atgcatttac gttgacacca tcgaatggtg
     2701 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     2761 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     2821 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     2881 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     2941 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     3001 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     3061 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     3121 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     3181 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     3241 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     3301 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     3361 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     3421 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     3481 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     3541 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     3601 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     3661 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     3721 cgtgtcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     3781 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3841 gtgagcgcaa cgcaattaat gtaagttagc gcgaattgat ctggtttgac agcttatcat
     3901 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     3961 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     4021 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     4081 aattaatcat ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
     4141 aaacagacca tgtccagaga ccattaatgc agctggcacg acaggtttcc cgactggaaa
     4201 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4261 ttacacttta tgcttccgga tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4321 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
     4381 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
     4441 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4501 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4561 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     4621 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4681 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4741 tggtctctag cggttaagag aagattttca gcctgataca gattaaatca gaacgcagaa
     4801 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     4861 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtat ccccatgcga
     4921 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     4981 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     5041 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     5101 gccaggcatc aaattaagca gaaggccatc ctgacggatg gccttttctg gctggatgat
     5161 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc
     5221 cccccccccc ctgcaggtcg atctggtaac cccagcgcgg ttgctttgtt cgatccgctg
     5281 gccggtaaaa ccccaagtta tctatctggg agcttatctt tgggcctatc gcagcaatag
     5341 cctggatgcg gggccgccga tcgtggtgtt cgactatcaa accggccgca gtggtcaaca
     5401 tgccgccgag tttcttcagc actggccagc caatggacca accaatggac caaccaatgg
     5461 accaaccaat gtaccaacca atggaccaac caatggacca accaatgggc aaggcacctt
     5521 gatggtcgat gactacagcg ggtacaaagc cctgttccgt gcgggcgtga ccgagcttgg
     5581 ctgctgggcc catgcccggc gcaagttctt tgatctgcac aaggccagca atcatccggt
     5641 tgcgcacgag gcactgtcgc ggattgcccg gttgtatctt gcagaagaag aggcccaagg
     5701 catggacatt gcggggcgga tgcaacaccg tgccctgcac agcctgccgc agttgcagga
     5761 catgcaccag tg
//
