LOCUS       sequence-449926-        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: pJB74
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(76..95)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             87..902
                     /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             87..902
                     /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(160..179)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1046..1068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1146..1223
                     /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             1224..2306
                     /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(1243..1262)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2538..2543,2544..2560,2561..2567)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2575..2591
                     /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     2580..2602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5248..5265)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5418..5504
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5596..5623
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      6446..7034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6935..6954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
       61 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga
      121 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
      181 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
      241 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
      301 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
      361 atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag
      421 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
      481 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcga ctcgctcagg
      541 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
      601 gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt
      661 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
      721 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
      781 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
      841 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
      901 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
      961 gcggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc
     1021 ccttacagac aagctgtgac cgcctccggg agctgcatgt gtcagaggtt ttcaccgtca
     1081 tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg gcatgcattt
     1141 acgttgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     1201 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     1261 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     1321 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     1381 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     1441 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     1501 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     1561 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     1621 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     1681 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     1741 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     1801 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     1861 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     1921 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     1981 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     2041 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     2101 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     2161 tgaagggcaa tcagctgttg cccgtgtcac tggtgaaaag aaaaaccacc ctggcgccca
     2221 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2281 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gcgcgaattg
     2341 atctggtttg acagcttatc atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     2401 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     2461 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     2521 tattctgaaa tgagctgttg acaattaatc atccggctcg tataatgtgt ggaattgtga
     2581 gcggataaca atttcacaca ggaaacagac catgtcccct tcacagtcag gaatgaatcc
     2641 tgccgacctg agcggactct ctggcaagga actggcacgc gcacgccgcg ctgcactatc
     2701 caagcaaggg aaagcagctg tttctaataa aacggctagc gtaaaccgta gcactaaaca
     2761 ggcggcatct tcgatcaata caaatcaggt gcgctcttct gtaaatgaag tgcccactga
     2821 ttaccaaatg gcggatcaat tgtgctctac gattgatcat gctgactttg gtaccgaaag
     2881 caatcgcgtt agagatctct gccgtcaacg cagagaggca ctatcaacta tcggtaaaaa
     2941 agcggttaaa accaacggca agccgtcggg tcgcgttcga ccacagcaat cagtggttca
     3001 caacgacgca atgatcgaaa atgccggtga tactaaccaa tcatcgtcca cttcattaaa
     3061 taatgaactt tccgaaatct gctccatagc agacgacatg ccggagcgtt ttggttcaca
     3121 agccaaaacc gtccgtgata tctgccgtgc acgccgtcaa gcgctctctg agcgtggaac
     3181 tcgcgccgtg ccgccaaagc cgcaatctca aggtggtcca ggacgcaatg gctatcaaat
     3241 tgatggatac ctagataccg cacttcatgg ccgcgatgcc gccaagcgcc accgtgaaat
     3301 gctctgtcaa tacggccgcg gcacagcacc ttcctgcaag ccaacaggcc gtgtcaaaaa
     3361 ttctgtacag tcgggcaacg cagcgccaaa aaaggttgaa accggtcaca ccctatccgg
     3421 cggatctgtt acgggcacgc aagtggatcg taaatctcat gtgactggaa acgagccggg
     3481 cacttgccga gcagtcacgg gcaccgaggc agtaggtact gagcaattca cctctttttg
     3541 caataccagc cccaagccaa atgcgacgaa ggtcaatgtg accacaacgg ctcgtggtcg
     3601 ccctgttagc ggtacggaag tttcacggac cgagaaggta actggcaacg aatccggtgt
     3661 ctgccgtaac gttaccggca ccgaagcgat gagtaatgaa gctcactttt ctctatgtgg
     3721 aacagccgca aagccttcac aagcggataa agtcatgttc ggcgccacag cacgaacgca
     3781 tcaagtggtc agtggcagtg atgaattcag gccctcttct gttacgggta acgaatcggg
     3841 tgcaaaacgc acaattaccg gctcgcaggc cgcagacgaa ggtcttgcgc gactcacgat
     3901 caacggagca cctgcaaaag tagccagaac ccacaccttt gcgggctctg acgttaccgg
     3961 cacggaaatc ggtcgctcta ctcgcgtaac tggtgatgaa agcggttcgt gtcgttcaat
     4021 ctcaggcacc gaggccctca gtaacgagca attccaatcc ttttgtgaca caaaacctca
     4081 acgcagcccg ttcaaggttg gccaagatcg cacgaacaag ggtcagtctg tgactggtaa
     4141 cttggttgat cgttccgaac tggttacagg taacgaacca ggttcatgct ctcgggttac
     4201 aggctctcag gcaggccaaa gcaaaatctg cggtggtggc gtgggaaaag tgcgctcaat
     4261 gcgcaccctt cgcggcacct cagtatctgg ccaacagcta gatcatgccc caaagatgtc
     4321 cggtgacgag cgcggcgggt gcatgcccgt caccggtaat gagtactacg gtcgtgaaca
     4381 tttcgaaccg ttttgtacga gcaccccaga gcccgaagct caatcaactg aacaatcatt
     4441 gacctgtgaa ggacaaatta ttagcggcac ttcagttgac gccagtgatt tggtcacagg
     4501 aaatgaaatc ggtgaacagc aactcatcag cggtgacgcc tatgttggcg cgcagcagac
     4561 aggttgcctt cccactagtc cacgcttcaa ccaaactggc aatgttcagt caatgggttt
     4621 taagaacacc aatcagccag aacaaaactt tgcaccaggt gaagtaatgc ctactgactt
     4681 tagtattcaa accccagctc gctcggctca gaatcgcatt acaggtaacg acattgcgcc
     4741 ctcaggtcgc attacaggcc ctggtatgct ggcaaccggc ttgattacag gaacccccga
     4801 attcaggcac gctgcgcgcg agttggttgg ttctccacaa cccatggcaa tggccatggc
     4861 caaccgtaat aaggctgctc aagcacctgt tgtgcagcca gaagtggttg caactcagga
     4921 aaagcctgag ttggtatgtg caccaagaag cgatcaaatg gatcgtgtga gtggcgaagg
     4981 caaagaacgt tgccacatca ctggcgatga ctggtcagtt aacaagcaca tcaccggtac
     5041 agccggtcaa tgggcgagtg gtcgcaaccc ttccatgcgc ggtaatgcgc gtgtggtcga
     5101 aaccagcgcg tttgccaatc gcaatgtgcc aaaacctgaa aagccgggct ccaagatcac
     5161 gggcagtagt ggtaatgaca cccaaggtag tctgatcact tactccggcg gcgcgcgcgg
     5221 ttgaagcggt taagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     5281 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     5341 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtatcccc atgcgagagt
     5401 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     5461 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     5521 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     5581 ggcatcaaat taagcagaag gccatcctga cggatggcct tttctggctg gatgatgggg
     5641 cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct cagcgccccc
     5701 ccccccctgc aggtcgatct ggtaacccca gcgcggttgc tttgttcgat ccgctggccg
     5761 gtaaaacccc aagttatcta tctgggagct tatctttggg cctatcgcag caatagcctg
     5821 gatgcggggc cgccgatcgt ggtgttcgac tatcaaaccg gccgcagtgg tcaacatgcc
     5881 gccgagtttc ttcagcactg gccagccaat ggaccaacca atggaccaac caatggacca
     5941 accaatgtac caaccaatgg accaaccaat ggaccaacca atgggcaagg caccttgatg
     6001 gtcgatgact acagcgggta caaagccctg ttccgtgcgg gcgtgaccga gcttggctgc
     6061 tgggcccatg cccggcgcaa gttctttgat ctgcacaagg ccagcaatca tccggttgcg
     6121 cacgaggcac tgtcgcggat tgcccggttg tatcttgcag aagaagaggc ccaaggcatg
     6181 gacattgcgg ggcggatgca acaccgtgcc ctgcacagcc tgccgcagtt gcaggacatg
     6241 caccagtggc tcagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg
     6301 gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     6361 cctatgccct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6421 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6481 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6541 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6601 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6661 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6721 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6781 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6841 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6901 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6961 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7021 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7081 cttttgctca catgttcttt cctgcgttag gacccaacgc tgcccgagat gcgccgcgtg
     7141 cggctgctgg agatggcgga cgcgatggat atgttctgcc acctgcgact caaacttctg
     7201 atctgggatg gcacgggggt gtggctctgt cagcgccggt tgcaccgggg gcatttttat
     7261 tggccaaagc ccggggatga tcggtttatc atgagtgccg aacactggca gtggctgacc
     7321 accggggttg actggcagag gctctgcgcc aagccacagg cccactggca agtgtgagat
     7381 ttttcaggat ttcgtcttga aaaatcagca catgcagcct ggggtttggt ataatcaacc
     7441 tcatgatttc aaacgcagaa accggtcatt catcccctga aacgccctcc gtggcgtggc
     7501 tgattgcgca actggaactg cgggattctg agttggcaaa acttgatgcg gcactggtgc
     7561 aacgcaatca agacatccgc catttaaccc aaaatcttca tgccaaagac atcaagatca
     7621 aggcactcac ccatgaactg gcgtattacc gccgtatccg cttcggggcc aaaaccgagg
     7681 ccatgagtgc cgagcaactg cacctgttcg aagacgatct ggccgaggta gtaagcttag
     7741 atcgacctgc aggggggggg ggaaagccac gttgtgtctc aaaatc
//
