LOCUS       sequence-141881-        7672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 hypothetical acetic acid resistance 
            protein B, cleavable N-terminal NusA fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  AUTHORS   Kappock
  TITLE     Additional Acetobacter aceti 1023 acetic acid resistance (aar) gene 
            constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7672)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJK122
FEATURES             Location/Qualifiers
     source          1..7672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(582..1373,1374..1442))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1205..1224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1535..2123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2024..2043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2277..2294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2309..2451
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2377..2396)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2537..2559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2553..2744)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3316..4398)
                     /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     4360..4379
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4399..4476)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4682..4701
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4789..4808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4789..4807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4808..4832
                     /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)."
     RBS             4847..4869
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4877..6361
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             6377..6394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6404..6448
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             6464..6481
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6500..6514
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             7105..7137
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             7144..7161
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      7327..7413
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7505..7532
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(7586..7604)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7600..7647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
      481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
      721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
      781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
      841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
      901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
      961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1441 atactcttcc tttttcaatc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1501 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1561 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1621 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1681 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1741 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1801 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1861 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1921 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1981 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2041 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2101 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2161 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2221 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2281 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     2341 gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata
     2401 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     2461 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2521 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2581 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     2641 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     2701 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     2761 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     2821 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     2881 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     2941 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     3001 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     3061 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     3121 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     3181 aacgacagga gcacgatcat gctagtcatg ccccgcgccc accggaagga gctgactggg
     3241 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     3301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     3421 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     3481 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     3541 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc
     3601 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     3661 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     3721 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     3781 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     3841 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     3901 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     3961 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     4021 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     4081 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     4141 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     4201 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     4261 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     4321 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     4381 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     4441 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     4501 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     4561 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     4621 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     4681 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     4741 cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta atacgactca
     4801 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta
     4861 agaaggagat atacatatga acaaagaaat tttggctgta gttgaagccg tatccaatga
     4921 aaaggcgcta cctcgcgaga agattttcga agcattggaa agcgcgctgg cgacagcaac
     4981 aaagaaaaaa tatgaacaag agatcgacgt ccgcgtacag atcgatcgca aaagcggtga
     5041 ttttgacact ttccgtcgct ggttagttgt tgatgaagtc acccagccga ccaaggaaat
     5101 cacccttgaa gccgcacgtt atgaagatga aagcctgaac ctgggcgatt acgttgaaga
     5161 tcagattgag tctgttacct ttgaccgtat cactacccag acggcaaaac aggttatcgt
     5221 gcagaaagtg cgtgaagccg aacgtgcgat ggtggttgat cagttccgtg aacacgaagg
     5281 tgaaatcatc accggcgtgg tgaaaaaagt aaaccgcgac aacatctctc tggatctggg
     5341 caacaacgct gaagccgtga tcctgcgcga agatatgctg ccgcgtgaaa acttccgccc
     5401 tggcgaccgc gttcgtggcg tgctctattc cgttcgcccg gaagcgcgtg gcgcgcaact
     5461 gttcgtcact cgttccaagc cggaaatgct gatcgaactg ttccgtattg aagtgccaga
     5521 aatcggcgaa gaagtgattg aaattaaagc agcggctcgc gatccgggtt ctcgtgcgaa
     5581 aatcgcggtg aaaaccaacg ataaacgtat cgatccggta ggtgcttgcg taggtatgcg
     5641 tggcgcgcgt gttcaggcgg tgtctactga actgggtggc gagcgtatcg atatcgtcct
     5701 gtgggatgat aacccggcgc agttcgtgat taacgcaatg gcaccggcag acgttgcttc
     5761 tatcgtggtg gatgaagata aacacaccat ggacatcgcc gttgaagccg gtaatctggc
     5821 gcaggcgatt ggccgtaacg gtcagaacgt gcgtctggct tcgcaactga gcggttggga
     5881 actcaacgtg atgaccgttg acgacctgca agctaagcat caggcggaag cgcacgcagc
     5941 gatcgacacc ttcaccaaat atctcgacat cgacgaagac ttcgcgactg ttctggtaga
     6001 agaaggcttc tcgacgctgg aagaattggc ctatgtgccg atgaaagagc tgttggaaat
     6061 cgaaggcctt gatgagccga ccgttgaagc actgcgcgag cgtgctaaaa atgcactggc
     6121 caccattgca caggcccagg aagaaagcct cggtgataac aaaccggctg acgatctgct
     6181 gaaccttgaa ggggtagatc gtgatttggc attcaaactg gccgcccgtg gcgtttgtac
     6241 gctggaagat ctcgccgaac agggcattga tgatctggct gatatcgaag ggttgaccga
     6301 cgaaaaagcc ggagcactga ttatggctgc ccgtaatatt tgctggttcg gtgacgaagc
     6361 gactagtggt tctggtcatc accatcacca tcactccgcg ggtaaagaaa ccgctgctgc
     6421 gaaatttgaa cgccagcaca tggactcgcc accgccaact ggtctggtcc cccggggcag
     6481 cgcgggttct ggtacgattg atgacgacga caaaatgctg ggtaaaccgc agttctttgg
     6541 tctgcgcgtt gctcgcatgt gtgcgaacgc atgtttcggt caccgtactt tcgcatctgt
     6601 tctgtatctg gccacctttc gttaccgtac cttccagaag cagcagcagc gtctcgaacg
     6661 cattccggat ccagctcact ggccagcgtg cctgaccgac gcaccggctc gtaaataccg
     6721 tggctatgca cctccgtccg cgctgtgtgg tctgaatcgt gaacgtgctg agttcgcgcc
     6781 gactaaactg caggcatggc gtcgcaaata cctgcataac atccgtgcat gggaaggcgg
     6841 tatgccggtg ccgccgcaac gtctgccgga atctgcggta tgcgcactgc ccgggtacat
     6901 caaatggtac cgcgcgcagt gccgtgcact gccggccagc gctcactccg ctgttccgcc
     6961 gctggcactg cgtgcttgag aattctgtac agagtccggg agctcgtgga tccgaattct
     7021 gtacaggcgc gcctgcagga cgtcgacggt accatcgata cgcgttcgaa gcttgcggcc
     7081 gcacagctgt atacacgtgc aagccagcca gaactcgctc ctgaagaccc agaggatctc
     7141 gagcaccacc accaccacca ctaatgttaa ttaagttggg cgttcctagg ctgataaaac
     7201 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     7261 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     7321 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     7381 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     7441 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     7501 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     7561 tttctaaata cattcaaata tgtatccgct gagcaataac tagcataacc ccttggggcc
     7621 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg at
//
