LOCUS       sequence-449915-        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: pJB44
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..94)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             86..901
                     /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             86..901
                     /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(159..178)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1045..1067
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1145..1222
                     /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             1223..2305
                     /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(1242..1261)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2537..2542,2543..2559,2560..2566)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2574..2590
                     /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     2579..2601
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5247..5264)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5417..5503
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5595..5622
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      6445..7033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6934..6953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
       61 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctcgag
      121 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
      181 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
      241 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
      301 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
      361 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
      421 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
      481 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgac tcgctcaggc
      541 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
      601 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
      661 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
      721 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
      781 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
      841 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
      901 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
      961 cggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc
     1021 cttacagaca agctgtgacc gcctccggga gctgcatgtg tcagaggttt tcaccgtcat
     1081 caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta
     1141 cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     1201 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     1261 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1321 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     1381 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     1441 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     1501 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     1561 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     1621 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     1681 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     1741 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     1801 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     1861 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     1921 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     1981 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     2041 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     2101 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     2161 gaagggcaat cagctgttgc ccgtgtcact ggtgaaaaga aaaaccaccc tggcgcccaa
     2221 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2281 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag cgcgaattga
     2341 tctggtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     2401 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     2461 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     2521 attctgaaat gagctgttga caattaatca tccggctcgt ataatgtgtg gaattgtgag
     2581 cggataacaa tttcacacag gaaacagacc atgtcccctt cacagtcagg aatgaatcct
     2641 gccgacctga gcggactctc tggcaaggaa ctggcacgcg cacgccgcgc tgcactatcc
     2701 aagcaaggga aagcagctgt ttctaataaa acggctagcg taaaccgtag cactaaacag
     2761 gcggcatctt cgatcaatac aaatcaggtg cgctcttctg taaatgaagt gcccactgat
     2821 taccaaatgg cggatcaatt gtgctctacg attgatcatg ctgactttgg taccgaaagc
     2881 aatcgcgtta gagatctctg ccgtcaacgc agagaggcac tatcaactat cggtaaaaaa
     2941 gcggttaaaa ccaacggcaa gccgtcgggt cgcgttcgac cacagcaatc agtggttcac
     3001 aacgacgcaa tgatcgaaaa tgccggtgat actaaccaat catcgtccac ttcattaaat
     3061 aatgaacttt ccgaaatctg ctccatagca gacgacatgc cggagcgttt tggttcacaa
     3121 gccaaaaccg tccgtgatat ctgccgtgca cgccgtcaag cgctctctga gcgtggaact
     3181 cgcgccgtgc cgccaaagcc gcaatctcaa ggtggtccag gacgcaatgg ctatcaaatt
     3241 gatggatacc tagataccgc acttcatggc cgcgatgccg ccaagcgcca ccgtgaaatg
     3301 ctctgtcaat acggccgcgg cacagcacct tcctgcaagc caacaggccg tgtcaaaaat
     3361 tctgtacagt cgggcaacgc agcgccaaaa aaggttgaaa ccggtcacac cctatccggc
     3421 ggatctgtta cgggcacgca agtggatcgt aaatctcatg tgactggaaa cgagccgggc
     3481 actagccgag cagtcacggg caccgagtac gtaggtactg agcaattcac ctcttttagc
     3541 aataccagcc ccaagccaaa tgcgacgaag gtcaatgtga ccacaacggc tcgtggtcgc
     3601 cctgttagcg gtacggaagt ttcacggacc gagaaggtaa ctggcaacga atccggtgtc
     3661 tgccgtaacg ttaccggcac cgaatacatg agtaatgaag ctcacttttc tctatgtgga
     3721 acagccgcaa agccttcaca agcggataaa gtcatgttcg gcgccacagc acgaacgcat
     3781 caagtggtca gtggcagtga tgaattcagg ccctcttctg ttacgggtaa cgaatcgggt
     3841 gcaaaacgca caattaccgg ctcgcagtac gcagacgaag gtcttgcgcg actcacgatc
     3901 aacggagcac ctgcaaaagt agccagaacc cacacctttg cgggctctga cgttaccggc
     3961 acggaaatcg gtcgctctac tcgcgtaact ggtgatgaaa gcggttcgtg tcgttcaatc
     4021 tcaggcaccg agtatctcag taacgagcaa ttccaatcct tttgtgacac aaaacctcaa
     4081 cgcagcccgt tcaaggttgg ccaagatcgc acgaacaagg gtcagtctgt gactggtaac
     4141 ttggttgatc gttccgaact ggttacaggt aacgaaccag gttcatgctc tcgggttaca
     4201 ggctctcagt atggccaaag caaaatctgc ggtggtggcg tgggaaaagt gcgctcaatg
     4261 cgcacccttc gcggcacctc agtatctggc caacagctag atcatgcccc aaagatgtcc
     4321 ggtgacgagc gcggcgggtg catgcccgtc accggtaatg agtactacgg tcgtgaacat
     4381 ttcgaaccgt tttgtacgag caccccagag cccgaagctc aatcaactga acaatcattg
     4441 acctgtgaag gacaaattat tagcggcact tcagttgacg ccagtgattt ggtcacagga
     4501 aatgaaatcg gtgaacagca actcatcagc ggtgacgcct atgttggcgc gcagcagaca
     4561 ggttgccttc ccactagtcc acgcttcaac caaactggca atgttcagtc aatgggtttt
     4621 aagaacacca atcagccaga acaaaacttt gcaccaggtg aagtaatgcc tactgacttt
     4681 agtattcaaa ccccagctcg ctcggctcag aatcgcatta caggtaacga cattgcgccc
     4741 tcaggtcgca ttacaggccc tggtatgctg gcaaccggct tgattacagg aacccccgaa
     4801 ttcaggcacg ctgcgcgcga gttggttggt tctccacaac ccatggcaat ggccatggcc
     4861 aaccgtaata aggctgctca agcacctgtt gtgcagccag aagtggttgc aactcaggaa
     4921 aagcctgagt tggtatgtgc accaagaagc gatcaaatgg atcgtgtgag tggcgaaggc
     4981 aaagaacgtt gccacatcac tggcgatgac tggtcagtta acaagcacat caccggtaca
     5041 gccggtcaat gggcgagtgg tcgcaaccct tccatgcgcg gtaatgcgcg tgtggtcgaa
     5101 accagcgcgt ttgccaatcg caatgtgcca aaacctgaaa agccgggctc caagatcacg
     5161 ggcagtagtg gtaatgacac ccaaggtagt ctgatcactt actccggcgg cgcgcgcggt
     5221 tgaagcggtt aagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     5281 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     5341 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtatcccca tgcgagagta
     5401 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     5461 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     5521 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     5581 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt ttctggctgg atgatggggc
     5641 gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc agcgcccccc
     5701 cccccctgca ggtcgatctg gtaaccccag cgcggttgct ttgttcgatc cgctggccgg
     5761 taaaacccca agttatctat ctgggagctt atctttgggc ctatcgcagc aatagcctgg
     5821 atgcggggcc gccgatcgtg gtgttcgact atcaaaccgg ccgcagtggt caacatgccg
     5881 ccgagtttct tcagcactgg ccagccaatg gaccaaccaa tggaccaacc aatggaccaa
     5941 ccaatgtacc aaccaatgga ccaaccaatg gaccaaccaa tgggcaaggc accttgatgg
     6001 tcgatgacta cagcgggtac aaagccctgt tccgtgcggg cgtgaccgag cttggctgct
     6061 gggcccatgc ccggcgcaag ttctttgatc tgcacaaggc cagcaatcat ccggttgcgc
     6121 acgaggcact gtcgcggatt gcccggttgt atcttgcaga agaagaggcc caaggcatgg
     6181 acattgcggg gcggatgcaa caccgtgccc tgcacagcct gccgcagttg caggacatgc
     6241 accagtggct cagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg
     6301 cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     6361 ctatgccctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6421 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6481 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6541 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6601 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6661 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6721 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6781 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6841 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6901 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6961 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7021 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7081 ttttgctcac atgttctttc ctgcgttagg acccaacgct gcccgagatg cgccgcgtgc
     7141 ggctgctgga gatggcggac gcgatggata tgttctgcca cctgcgactc aaacttctga
     7201 tctgggatgg cacgggggtg tggctctgtc agcgccggtt gcaccggggg catttttatt
     7261 ggccaaagcc cggggatgat cggtttatca tgagtgccga acactggcag tggctgacca
     7321 ccggggttga ctggcagagg ctctgcgcca agccacaggc ccactggcaa gtgtgagatt
     7381 tttcaggatt tcgtcttgaa aaatcagcac atgcagcctg gggtttggta taatcaacct
     7441 catgatttca aacgcagaaa ccggtcattc atcccctgaa acgccctccg tggcgtggct
     7501 gattgcgcaa ctggaactgc gggattctga gttggcaaaa cttgatgcgg cactggtgca
     7561 acgcaatcaa gacatccgcc atttaaccca aaatcttcat gccaaagaca tcaagatcaa
     7621 ggcactcacc catgaactgg cgtattaccg ccgtatccgc ttcggggcca aaaccgaggc
     7681 catgagtgcc gagcaactgc acctgttcga agacgatctg gccgaggtag taagcttaga
     7741 tcgacctgca gggggggggg gaaagccacg ttgtgtctca aaatct
//
