LOCUS       sequence-147673-        6375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6375)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I
  TITLE     Expression vectors for the engineering of genes and genomes in 
            Acinetobacter baylyi ADP1.
  JOURNAL   Appl Environ Microbiol. 2011 Oct 21.
  PUBMED    22020504
REFERENCE   2  (bases 1 to 6375)
  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: "Appl 
            Environ Microbiol. 2011 Oct 21."
COMMENT     Sequence Label: lacI-PT5-gusA-pBAV1k
FEATURES             Location/Qualifiers
     source          1..6375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      70..156
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1487..1498
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(1700..1794)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1810..2604)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    2810..2829
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        2856..2933
                     /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             2934..4016
                     /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(2953..2972)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4065..4070,4071..4088,4089..4095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4119
                     /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     4127..4143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4127..4143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4159..4176)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4159..4175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4168..4190)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4333..4351
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4375..4389,4390..4395,4396..4412,4413..4419)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    4395..4411
                     /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    4427..4443
                     /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             4466..4477
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4496..4513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4538..6343
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
                     VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
                     EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
                     FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
                     VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFL
                     INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
                     DEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
                     NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     misc_feature    join(6374..6375,1..19)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 ctagtagcgg ccgctgcagg cctcagggcc cgatcgatgc cgccgcttaa ttaattaatc
       61 cagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      121 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagtgtcat
      181 tttatttccc ccgtttcagc atcaagaacc tttgcataac ttgctctata tccacactga
      241 taattgccct caaaccataa tctaaaggcg ctagagtttg ttgaaacaat atcttttaca
      301 tcattcgtat ttaaaattcc aaactccgct cccctaaggc gaataaaagc cattaaatct
      361 tttgtattta ccaaattata gtcatccact atatctaaga gtaaattctt caattctctt
      421 ttttggcttt catcaagtgt tatatagcgg tcaatatcaa aatcattaat gttcaaaata
      481 tcttttttgt cgtatatatg tttattctta gcaatagcgt cctttgattc atgagtcaaa
      541 tattcatatg aacctttgat ataatcaagt atctcaacat gagcaactga actattcccc
      601 aattttcgct taatcttgtt cctaacgctt tctattgtta caggatttcg tgcaatatat
      661 ataacgtgat agtgtggttt tttatagtgc tttccatttc gtataacatc actactattc
      721 catgtatctt tatctttttt ttcgtccata tcgtgtaaag gactgacagc catagatacg
      781 cccaaactct ctaatttttc cttccaatca ttaggaattg agtcaggata taataaaaat
      841 ccaaaatttc tagctttagt atttttaata gccatgatat aattacctta tcaaaaacaa
      901 gtagcgaaaa ctcgtatcct tctaaaaacg cgagctttcg cttatttttt ttgttctgat
      961 tcctttcttg catattcttc tatagctaac gccgcaaccg cagattttga aaaacctttt
     1021 tgtttcgcca tatctgttaa ttttttatct tgctcttttg tcagagaaat cataactctt
     1081 tttttcgatt ctgaaatcac catttaaaaa actccaatca aataatttta taaagttagt
     1141 gtatcacttt gtaatcataa aaacaacaat aaagctactt aaatatagat ttataaaaaa
     1201 cgttggcgaa aacgttggcg attcgttggc gattgaaaaa ccccttaaac ccttgagcca
     1261 gttgggatag agcgtttttg gcacaaaaat tggcactcgg cacttaatgg ggggtcgtag
     1321 tacggaagca aaattcgctt cctttccccc catttttttc caaattccaa atttttttca
     1381 aaaattttcc agcgctaccg ctcggcaaaa ttgcaagcaa tttttaaaat caaacccatg
     1441 agggaatttc attccctcat actcccttga gcctcctcca accgaaatag aagggcgctg
     1501 cgcttattat ttcattcagt catcggcttt cataatctaa cagacaacat cttcgctgca
     1561 aagccacgct acgctcaagg gcttttacgc tacgataacg cctgttttaa cgattatgcc
     1621 gataactaaa cgaaataaac gctaaaacgt ctcagaaacg attttgagac gttttaataa
     1681 aaaatcgcct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     1741 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     1801 agctcggtac taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg
     1861 cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct
     1921 gatcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag
     1981 atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc
     2041 gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg
     2101 cggatcttta aatggagtgt cttcttccca gttttcgcaa tccacatcgg ccagatcgtt
     2161 attcagtaag taatccaatt cggctaagcg gccgtctaag ctattcgtat agggacaatc
     2221 cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga tagtcttttc
     2281 agggctttgt tcatcttcat acccttccga gcaaaggacg ccatcggcct cactcatgag
     2341 cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc
     2401 ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg
     2461 gctgtccgtc atttttaaat ataggatttc attttctccc accagcttat ataccttagc
     2521 aggagacatt ccttccgtat cttttacgca gcggtattct tcgatcagtt ttttcaattc
     2581 cggtgatatt ctcattttag ccatttatta tttccttcct cttttctaca gtatttaaag
     2641 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     2701 aaccttaaat acagaaaaca gccttttcaa agttgttttc aaagttggcg tataacatag
     2761 tatcgacgga gccgattttg aaaccacaat tatgatagaa tttgacgtcg aattcgcggc
     2821 cgcttctaga actatagcta gcatgcattt acgttgacac catcgaatgg tgcaaaacct
     2881 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     2941 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     3001 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     3061 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     3121 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     3181 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     3241 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     3301 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     3361 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     3421 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     3481 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     3541 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     3601 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     3661 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     3721 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     3781 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     3841 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     3901 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3961 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4021 aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4081 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4141 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4201 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4261 agaggcccgc accgatcgcc cttcccaaca gttgcctaag aaaccattat tatcatgaca
     4321 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gaaatcataa
     4381 aaaatttatt tgctttgtga gcggataaca attataatag attcaattgt gagcggataa
     4441 caatttcaca cagaattaat tcattaaaga ggagaaatta accatgggca gcagccatca
     4501 ccatcatcac cacagccagg atccggccct gagggcctta cgtcctgtag aaaccccaac
     4561 ccgtgaaatc aaaaaactcg acggcctgtg ggcattcagt ctggatcgcg aaaactgtgg
     4621 aattgatcag cgttggtggg aaagcgcgtt acaagaaagc cgggcaattg ctgtgccagg
     4681 cagttttaac gatcagttcg ccgatgcaga tattcgtaat tatgcgggca acgtctggta
     4741 tcagcgcgaa gtctttatac cgaaaggttg ggcaggccag cgtatcgtgc tgcgtttcga
     4801 tgcggtcact cattacggca aagtgtgggt caataatcag gaagtgatgg agcatcaggg
     4861 cggctatacg ccatttgaag ccgatgtcac gccgtatgtt attgccggga aaagtgtacg
     4921 tatcaccgtt tgtgtgaaca acgaactgaa ctggcagact atcccgccgg gaatggtgat
     4981 taccgacgaa aacggcaaga aaaagcagtc ttacttccat gatttcttta actatgccgg
     5041 gatccatcgc agcgtaatgc tctacaccac gccgaacacc tgggtggacg atatcaccgt
     5101 ggtgacgcat gtcgcgcaag actgtaacca cgcgtctgtt gactggcagg tggtggccaa
     5161 tggtgatgtc agcgttgaac tgcgtgatgc ggatcaacag gtggttgcaa ctggacaagg
     5221 cactagcggg actttgcaag tggtgaatcc gcacctctgg caaccgggtg aaggttatct
     5281 ctatgaactg tgcgtcacag ccaaaagcca gacagagtgt gatatctacc cgcttcgcgt
     5341 cggcatccgg tcagtggcag tgaagggcga acagttcctg attaaccaca aaccgttcta
     5401 ctttactggc tttggtcgtc atgaagatgc ggacttgcgt ggcaaaggat tcgataacgt
     5461 gctgatggtg cacgaccacg cattaatgga ctggattggg gccaactcct accgtacctc
     5521 gcattaccct tacgctgaag agatgctcga ctgggcagat gaacatggca tcgtggtgat
     5581 tgatgaaact gctgctgtcg gctttaacct ctctttaggc attggtttcg aagcgggcaa
     5641 caagccgaaa gaactgtaca gcgaagaggc agtcaacggg gaaactcagc aagcgcactt
     5701 acaggcgatt aaagagctga tagcgcgtga caaaaaccac ccaagcgtgg tgatgtggag
     5761 tattgccaac gaaccggata cccgtccgca aggtgcacgg gaatatttcg cgccactggc
     5821 ggaagcaacg cgtaaactcg acccgacgcg tccgatcacc tgcgtcaatg taatgttctg
     5881 cgacgctcac accgatacca tcagcgatct ctttgatgtg ctgtgcctga accgttatta
     5941 cggatggtat gtccaaagcg gcgatttgga aacggcagag aaggtactgg aaaaagaact
     6001 tctggcctgg caggagaaac tgcatcagcc gattatcatc accgaatacg gcgtggatac
     6061 gttagccggg ctgcactcaa tgtacaccga catgtggagt gaagagtatc agtgtgcatg
     6121 gctggatatg tatcaccgcg tctttgatcg cgtcagcgcc gtcgtcggtg aacaggtatg
     6181 gaatttcgcc gattttgcga cctcgcaagg catattgcgc gttggcggta acaagaaagg
     6241 gatcttcact cgcgaccgca aaccgaagtc ggcggctttt ctgctgcaaa aacgctggac
     6301 tggcatgaac ttcggtgaaa aaccgcagca gggaggcaaa caaggcctat gcggccggta
     6361 gaaaggagaa gctta
//
