LOCUS       sequence-449918-        7786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli and H. neapolitanus shuttle vector; integrates the gene of 
            interest and a kanamycin resistance marker into a neutral site on 
            the H. neapolitanus genome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7786)
  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 7786)
  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: pJB47
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(74..93)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             85..900
                     /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             85..900
                     /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(158..177)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1044..1066
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1144..1221
                     /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             1222..2304
                     /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(1241..1260)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2536..2541,2542..2558,2559..2565)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2573..2589
                     /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     2578..2600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5246..5263)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5416..5502
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5594..5621
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6933..6952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
       61 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctcgagg
      121 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
      181 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
      241 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
      301 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
      361 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
      421 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
      481 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgact cgctcaggcg
      541 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
      601 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
      661 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
      721 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
      781 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
      841 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
      901 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
      961 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccc
     1021 ttacagacaa gctgtgaccg cctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1081 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcatttac
     1141 gttgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     1201 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     1261 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     1321 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     1381 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     1441 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1501 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1561 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1621 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1681 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1741 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1801 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1861 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1921 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1981 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     2041 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     2101 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     2161 aagggcaatc agctgttgcc cgtgtcactg gtgaaaagaa aaaccaccct ggcgcccaat
     2221 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2281 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc gcgaattgat
     2341 ctggtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
     2401 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
     2461 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
     2521 ttctgaaatg agctgttgac aattaatcat ccggctcgta taatgtgtgg aattgtgagc
     2581 ggataacaat ttcacacagg aaacagacca tgtccccttc acagtcagga atgaatcctg
     2641 ccgacctgag cggactctct ggcaaggaac tggcacgcgc acgccgcgct gcactatcca
     2701 agcaagggaa agcagctgtt tctaataaaa cggctagcgt aaaccgtagc actaaacagg
     2761 cggcatcttc gatcaataca aatcaggtgc gctcttctgt aaatgaagtg cccactgatt
     2821 accaaatggc ggatcaattg tgctctacga ttgatcatgc tgactttggt accgaaagca
     2881 atcgcgttag agatctctgc cgtcaacgca gagaggcact atcaactatc ggtaaaaaag
     2941 cggttaaaac caacggcaag ccgtcgggtc gcgttcgacc acagcaatca gtggttcaca
     3001 acgacgcaat gatcgaaaat gccggtgata ctaaccaatc atcgtccact tcattaaata
     3061 atgaactttc cgaaatctgc tccatagcag acgacatgcc ggagcgtttt ggttcacaag
     3121 ccaaaaccgt ccgtgatatc tgccgtgcac gccgtcaagc gctctctgag cgtggaactc
     3181 gcgccgtgcc gccaaagccg caatctcaag gtggtccagg acgcaatggc tatcaaattg
     3241 atggatacct agataccgca cttcatggcc gcgatgccgc caagcgccac cgtgaaatgc
     3301 tctgtcaata cggccgcggc acagcacctt cctgcaagcc aacaggccgt gtcaaaaatt
     3361 ctgtacagtc gggcaacgca gcgccaaaaa aggttgaaac cggtcacacc ctatccggcg
     3421 gatctgttac gggcacgcaa gtggatcgta aatctcatgt gactggaaac gagccgggca
     3481 ctagccgagc agtcacgggc accgagtacg taggtactga gcaattcacc tcttttagca
     3541 ataccagccc caagccaaat gcgacgaagg tcaatgtgac cacaacggct cgtggtcgcc
     3601 ctgttagcgg tacggaagtt tcacggaccg agaaggtaac tggcaacgaa tccggtgtca
     3661 gccgtaacgt taccggcacc gaatacatga gtaatgaagc tcacttttct ctaagtggaa
     3721 cagccgcaaa gccttcacaa gcggataaag tcatgttcgg cgccacagca cgaacgcatc
     3781 aagtggtcag tggcagtgat gaattcaggc cctcttctgt tacgggtaac gaatcgggtg
     3841 caaaacgcac aattaccggc tcgcagtacg cagacgaagg tcttgcgcga ctcacgatca
     3901 acggagcacc tgcaaaagta gccagaaccc acacctttgc gggctctgac gttaccggca
     3961 cggaaatcgg tcgctctact cgcgtaactg gtgatgaaag cggttcgagt cgttcaatct
     4021 caggcaccga gtatctcagt aacgagcaat tccaatcctt tagtgacaca aaacctcaac
     4081 gcagcccgtt caaggttggc caagatcgca cgaacaaggg tcagtctgtg actggtaact
     4141 tggttgatcg ttccgaactg gttacaggta acgaaccagg ttcaagctct cgggttacag
     4201 gctctcagta tggccaaagc aaaatcagcg gtggtggcgt gggaaaagtg cgctcaatgc
     4261 gcacccttcg cggcacctca gtatctggcc aacagctaga tcatgcccca aagatgtccg
     4321 gtgacgagcg cggcgggtgc atgcccgtca ccggtaatga gtactacggt cgtgaacatt
     4381 tcgaaccgtt ttgtacgagc accccagagc ccgaagctca atcaactgaa caatcattga
     4441 cctgtgaagg acaaattatt agcggcactt cagttgacgc cagtgatttg gtcacaggaa
     4501 atgaaatcgg tgaacagcaa ctcatcagcg gtgacgccta tgttggcgcg cagcagacag
     4561 gttgccttcc cactagtcca cgcttcaacc aaactggcaa tgttcagtca atgggtttta
     4621 agaacaccaa tcagccagaa caaaactttg caccaggtga agtaatgcct actgacttta
     4681 gtattcaaac cccagctcgc tcggctcaga atcgcattac aggtaacgac attgcgccct
     4741 caggtcgcat tacaggccct ggtatgctgg caaccggctt gattacagga acccccgaat
     4801 tcaggcacgc tgcgcgcgag ttggttggtt ctccacaacc catggcaatg gccatggcca
     4861 accgtaataa ggctgctcaa gcacctgttg tgcagccaga agtggttgca actcaggaaa
     4921 agcctgagtt ggtatgtgca ccaagaagcg atcaaatgga tcgtgtgagt ggcgaaggca
     4981 aagaacgttg ccacatcact ggcgatgact ggtcagttaa caagcacatc accggtacag
     5041 ccggtcaatg ggcgagtggt cgcaaccctt ccatgcgcgg taatgcgcgt gtggtcgaaa
     5101 ccagcgcgtt tgccaatcgc aatgtgccaa aacctgaaaa gccgggctcc aagatcacgg
     5161 gcagtagtgg taatgacacc caaggtagtc tgatcactta ctccggcggc gcgcgcggtt
     5221 gaagcggtta agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     5281 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     5341 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtatccccat gcgagagtag
     5401 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5461 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     5521 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     5581 catcaaatta agcagaaggc catcctgacg gatggccttt tctggctgga tgatggggcg
     5641 attcaggcct ggtatgagtc agcaacacct tcttcacgag gcagacctca gcgccccccc
     5701 ccccctgcag gtcgatctgg taaccccagc gcggttgctt tgttcgatcc gctggccggt
     5761 aaaaccccaa gttatctatc tgggagctta tctttgggcc tatcgcagca atagcctgga
     5821 tgcggggccg ccgatcgtgg tgttcgacta tcaaaccggc cgcagtggtc aacatgccgc
     5881 cgagtttctt cagcactggc cagccaatgg accaaccaat ggaccaacca atggaccaac
     5941 caatgtacca accaatggac caaccaatgg accaaccaat gggcaaggca ccttgatggt
     6001 cgatgactac agcgggtaca aagccctgtt ccgtgcgggc gtgaccgagc ttggctgctg
     6061 ggcccatgcc cggcgcaagt tctttgatct gcacaaggcc agcaatcatc cggttgcgca
     6121 cgaggcactg tcgcggattg cccggttgta tcttgcagaa gaagaggccc aaggcatgga
     6181 cattgcgggg cggatgcaac accgtgccct gcacagcctg ccgcagttgc aggacatgca
     6241 ccagtggctc agaaattgca tcaacgcata tagcgctagc agcacgccat agtgactggc
     6301 gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
     6361 tatgccctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgttctttcc tgcgttagga cccaacgctg cccgagatgc gccgcgtgcg
     7141 gctgctggag atggcggacg cgatggatat gttctgccac ctgcgactca aacttctgat
     7201 ctgggatggc acgggggtgt ggctctgtca gcgccggttg caccgggggc atttttattg
     7261 gccaaagccc ggggatgatc ggtttatcat gagtgccgaa cactggcagt ggctgaccac
     7321 cggggttgac tggcagaggc tctgcgccaa gccacaggcc cactggcaag tgtgagattt
     7381 ttcaggattt cgtcttgaaa aatcagcaca tgcagcctgg ggtttggtat aatcaacctc
     7441 atgatttcaa acgcagaaac cggtcattca tcccctgaaa cgccctccgt ggcgtggctg
     7501 attgcgcaac tggaactgcg ggattctgag ttggcaaaac ttgatgcggc actggtgcaa
     7561 cgcaatcaag acatccgcca tttaacccaa aatcttcatg ccaaagacat caagatcaag
     7621 gcactcaccc atgaactggc gtattaccgc cgtatccgct tcggggccaa aaccgaggcc
     7681 atgagtgccg agcaactgca cctgttcgaa gacgatctgg ccgaggtagt aagcttagat
     7741 cgacctgcag gggggggggg aaagccacgt tgtgtctcaa aatctc
//
