LOCUS       Exported                9106 bp DNA     circular SYN 01-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pKLAC2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9106)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          989..1691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    257..354
                     /label=Multiple Cloning Site
                     /note="A synthetic DNA sequence containing multiple
                     restriction enzyme recognition sites, used for inserting 
                     foreign DNA fragments in molecular cloning experiments.; 
                     id: 4352484"
     terminator      371..988
                     /label=LAC4 terminator
                     /note="transcription terminator for Kluyveromyces lactis 
                     LAC4"
     promoter        989..1691
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1712..3358
                     /codon_start=1
                     /gene="amdS"
                     /product="acetamidase from Aspergillus nidulans"
                     /label=amdS
                     /note="confers ability to process acetamide for selection 
                     in fungal cells"
                     /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL
                     SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA
                     RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK
                     AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG
                     TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT
                     KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE
                     TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL
                     NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF
                     RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV
                     AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP"
     promoter        3994..4650
                     /label=LAC4 promoter (5')
                     /note="5' portion of the Kluyveromyces lactis LAC4 
                     promoter"
     promoter        complement(4667..4685)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4704..4826
                     /label=lac operon promoter
                     /note="Promoter region that regulates transcription of the 
                     lactose operon genes, including lacZ, controlling bacterial
                     metabolism of lactose in response to environmental 
                     conditions; id: 3799053"
     rep_origin      4857..5859
                     /label=pUC19 replication origin
                     /note="Origin of replication derived from pUC19 plasmid, 
                     containing the ColE1 origin sequence that enables 
                     autonomous DNA replication in bacterial hosts.; id: 
                     4370962"
     CDS             complement(5860..6720)
                     /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"
     promoter        6721..6851
                     /label=AmpR promoter
                     /note="Regulatory region that initiates transcription of
                     the ampicillin resistance gene in bacterial plasmids; id: 
                     3672594"
     rep_origin      6852..7307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7448..7464
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7478..9006
                     /label=LAC4 promoter (3')
                     /note="3' portion of a modified Kluyveromyces lactis LAC4 
                     promoter"
     protein_bind    9012..9028
                     /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)."
ORIGIN
        1 aagcttgaaa aaaatgaaat tctctactat attagccgca tctactgctt taatttccgt
       61 tgttatggct gctccagttt ctaccgaaac tgacatcgac gatcttccaa tatcggttcc
      121 agaagaagcc ttgattggat tcattgactt aaccggggat gaagtttcct tgttgcctgt
      181 taataacgga acccacactg gtattctatt cttaaacacc accatcgctg aagctgcttt
      241 cgctgacaag gatgatctcg agaaaagaga ggctgaagct agaagagctc atatgtccat
      301 gggcggccgc gatatcgtcg acggatccga attccctgca ggtaattaaa taaaggcctt
      361 gaatcgagaa tttatactta gataagtatg tacttacagg tatatttcta tgagatactg
      421 atgtatacat gcatgataat atttaaacgg ttattagtgc cgattgtctt gtgcgataat
      481 gacgttccta tcaaagcaat acacttacca cctattacat gggccaagaa aatattttcg
      541 aacttgttta gaatattagc acagagtata tgatgatatc cgttagatta tgcatgattc
      601 attcctacaa ctttttcgta gcataaggat taattacttg gatgccaata aaaaaaaaaa
      661 catcgagaaa atttcagcat gctcagaaac aattgcagtg tatcaaagta aaaaaaagat
      721 tttcactaca tgttcctttt gaagaaagaa aatcatggaa cattagattt acaaaaattt
      781 aaccaccgct gattaacgat tagaccgtta agcgcacaac aggttattag tacagagaaa
      841 gcattctgtg gtgttgcccc ggactttctt ttgcgacata ggtaaatcga ataccatcat
      901 actatctttt ccaatgactc cctaaagaaa gactcttctt cgatgttgta tacgttggag
      961 catagggcaa gaattgtggc ttgagatctt gattgtatgc ttggtatagc ttgaaatatt
     1021 gtgcagaaaa agaaacaagg aagaaaggga acgagaacaa tgacgaggaa acaaaagatt
     1081 aataattgca ggtctattta tacttgatag caagacagca aacttttttt tatttcaaat
     1141 tcaagtaact ggaaggaagg ccgtataccg ttgctcatta gagagtagtg tgcgtgaatg
     1201 aaggaaggaa aaagtttcgt gtgcttcgag atacccctca tcagctctgg aacaacgaca
     1261 tctgttggtg ctgtctttgt cgttaatttt ttcctttagt gtcttccatc atttttttgt
     1321 cattgcggat atggtgagac aacaacgggg gagagagaaa agaaaaaaaa agaaaagaag
     1381 ttgcatgcgc ctattattac ttcaatagat ggcaaatgga aaaagggtag tgaaacttcg
     1441 atatgatgat ggctatcaag tctagggcta cagtattagt tcgttatgta ccaccatcaa
     1501 tgaggcagtg taattggtgt agtcttgttt agcccattat gtcttgtctg gtatctgttc
     1561 tattgtatat ctcccctccg ccacctacat gttagggaga ccaacgaagg tattatagga
     1621 atcccgatgt atgggtttgg ttgccagaaa agaggaagtc catattgtac acccggaaac
     1681 aacaaaagga tggaattccg gatccgccac catgcctcaa tcctgggaag aactggccgc
     1741 tgataagcgc gcccgcctcg caaaaaccat ccctgatgaa tggaaagtcc agacgctgcc
     1801 tgcggaagac agcgttattg atttcccaaa gaaatcgggg atcctttcag aggccgaact
     1861 gaagatcaca gaggcttccg ctgcggatct tgtgtccaag ctggcggccg gagagttgac
     1921 ctcggtggaa gttacgctag cattctgtaa acgggcagca atcgcccagc agttaacaaa
     1981 ctgcgcccac gagttcttcc ctgacgccgc tctcgcgcag gcaagggaac tcgatgaata
     2041 ctacgcaaag cacaagagac ccgttggtcc actccatggc ctccccatct ctctcaaaga
     2101 ccagcttcga gtcaagggct acgaaacatc aatgggctac atctcatggc taaacaagta
     2161 cgacgaaggg gactcggttc tgacaaccat gctccgcaaa gccggtgccg tcttctacgt
     2221 caagacctct gtcccgcaga ccctgatggt ctgcgagaca gtcaacaaca tcatcgggcg
     2281 caccgtcaac ccacgcaaca agaactggtc gtgcggcggc agttctggtg gtgagggtgc
     2341 gatcgttggg attcgtggtg gcgtcatcgg tgtaggaacg gatatcggtg gctcgattcg
     2401 agtgccggcc gcgttcaact tcctgtacgg tctaaggccg agtcatgggc ggctgccgta
     2461 tgcaaagatg gcgaacagca tggagggtca ggagacggtg cacagcgttg tcgggccgat
     2521 tacgcactct gttgaggacc tccgcctctt caccaaatcc gtcctcggtc aggagccatg
     2581 gaaatacgac tccaaggtca tccccatgcc ctggcgccag tccgagtcgg acattattgc
     2641 ctccaagatc aagaacggcg ggctcaatat cggctactac aacttcgacg gcaatgtcct
     2701 tccacaccct cctatcctgc gcggcgtgga aactaccgtc gccgcactcg ccaaagccgg
     2761 tcacaccgtg accccgtgga cgccatacaa gcacgatttc ggccacgatc tcatctccca
     2821 tatctacgcg gctgacggca gcgccgacgt aatgcgcgat atcagtgcat ccggcgagcc
     2881 ggcgattcca aatatcaaag acctactgaa cccgaacatc aaagctgtta acatgaacga
     2941 gctctgggac acgcatctcc agaagtggaa ttaccagatg gagtaccttg agaaatggcg
     3001 ggaggctgaa gaaaaggccg ggaaggaact ggacgccatc atcgcgccga ttacgcctac
     3061 cgctgcggta cggcatgacc agttccggta ctatgggtat gcctctgtga tcaacctgct
     3121 ggatttcacg agcgtggttg ttccggttac ctttgcggat aagaacatcg ataagaagaa
     3181 tgagagtttc aaggcggtta gtgagcttga tgccctcgtg caggaagagt atgatccgga
     3241 ggcgtaccat ggggcaccgg ttgcagtgca ggttatcgga cggagactca gtgaagagag
     3301 gacgttggcg attgcagagg aagtggggaa gttgctggga aatgtggtga ctccatagcc
     3361 cggggggggc tcgatcccct cgcgagttgg ttcagctgct gcctgaggct ggacgacctc
     3421 gcggagttct accggcagtg caaatccgtc ggcatccagg aaaccagcag cggctatccg
     3481 cgcatccatg cccccgaact gcaggagtgg ggaggcacga tggccgcttt ggtcgatcta
     3541 gattacgtgg aagaaaggta gtaaaagtag tagtataagt agtaaaaaga ggtaaaaaga
     3601 gaaaaccggc tacatactag agaagcacgt acacaaaaac tcataggcac ttcatcatac
     3661 gacagtttct tgatgcatta taatagtgta ttagatattt tcagaaatat gcatagaacc
     3721 tcctcttgcc tttacttttt atacatagaa cattggcaga tttacttaca ctactttgtt
     3781 tctacgccat ttcttttgtt ttcaacactt agacaagttg ttgagaaccg gactactaaa
     3841 aagcaatgtt cccactgaaa atcatgtacc tgcaggataa taacccccta attctgcatc
     3901 gatccagtat gttttttttt ctctactcat ttttacctga agatagagct tctaaaacaa
     3961 aaaaaatcag tgattacatg catattgtgt gttctagtaa ccaaaggaaa ggaacagata
     4021 gataaaattc cgagactgtc aaattaggtt tttttctttt tttttggcgg gagtcagtgg
     4081 gccgaaatat gttcttggcc tagaacttaa tctggtttga tcatgccaat acttgcctga
     4141 gtgcccgact ttttgcccac cctcttgcct tctgtcatcc ttcaaaaccc acctgttttc
     4201 cagccgtatc ttcgctcgca tctacacata ctgtgccata tcttgtgtgt agccggacgt
     4261 gactatgacc aaaaacaaac aaggagaact gttcgccgat ttgtaacact cctgcatcca
     4321 tccaagtggg tatgcgctat gcaatgttaa gctaggtcag gtcagaccag gtccaaggac
     4381 agcaacttga ctgtatgcaa cctttaccat ctttgcacag aacatacttg tagctagcta
     4441 gttacactta tggaccgaaa aggcacccca ccatgtctgt ccggctttag agtacggccg
     4501 cagaccgctg atttgccttg ccaagcagta gtcacaatgc atcgcatgag cacacgggca
     4561 cgggcacggg cacaggaacc attggcaaaa ataccagata cactataccg acgtatatca
     4621 agcccaagtt taaaattcct aaatttccgc ggctactttt caattcccta tagtgagtcg
     4681 tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4741 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4801 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4861 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4921 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4981 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5041 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5281 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5521 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5581 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5941 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6001 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6061 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6121 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6181 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6241 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6301 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6361 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6421 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6481 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6541 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6601 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6661 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6721 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6781 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6841 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6901 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6961 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7021 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7081 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7141 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7201 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7261 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     7321 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7381 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     7441 cgacgttgta aaacgacggc cagtgccaag ctcccgcggg gatcgactca taaaatagta
     7501 accttctaat gcgtatctat tgactaccaa ccattagtgt ggttgcagaa ggcggaattc
     7561 tcccttcttc gaattcagct tgctttttca ttttttattt tccatttttc agtttttgtt
     7621 tgtgtcgaat ttagccagtt gcttctccaa gatgaaaaaa acccctgcgc agtttctgtg
     7681 ctgcaagatc ctaatcgact tttccacccc ccacaaaagt aaatgttctt ttgttacatt
     7741 cgcgtgggta gctagctccc cgaatcttca aaggacttag ggactgcact acatcagagt
     7801 gtgttcacct ggtttgctgc ctggtttgaa agaaaagagc agggaactcg cgggttcccg
     7861 gcgaataatc atgcgatagt cctttggcct tccaagtcgc atgtagagta gacaacagac
     7921 agggagggca ggaaggatct ttcactgaga tcctgtatct tgttgggtaa gtcggatgaa
     7981 aggggaatcg tatgagattg gagaggatgc ggaagaggta acgccttttg ttaacttgtt
     8041 taattattat ggggcaggcg agagggggag gaatgtatgt gtgtgaggcg ggcgagacgg
     8101 agccatccag gccaggtaga aatagagaaa gccgaatgtt agacaatatg gcagcgtagt
     8161 agagtaggta ggtaggcaag tactgctagc aaagaggaga agggtaagct cactcttcgc
     8221 attccacacc gttagtgtgt cagtttgaac aaaaaaacaa tcatcatacc aattgatgga
     8281 ctgtggactg gcttttggaa cggcttttcg gactgcgatt attcgtgagg aatcaaggta
     8341 ggaatttggt catatttacg gacaacagtg ggtgattccc atatggagta ggaaaacgag
     8401 atcatggtat cctcagatat gttgcggaat tctgttcacc gcaaagttca gggtgctctg
     8461 gtgggtttcg gttggtcttt gctttgcttc tcccttgtct tgcatgttaa taatagccta
     8521 gcctgtgagc cgaaacttag ggtaggctta gtgttggaac gtacatatgt atcacgttga
     8581 cttggtttaa ccaggcgacc tggtagccag ccatacccac acacgttttt tgtatcttca
     8641 gtatagttgt gaaaagtgta gcggaaattt gtggtccgag caacagcgtc tttttctagt
     8701 agtgcggtcg gttacttggt tgacattggt atttggactt tgttgctaca ccattcacta
     8761 cttgaagtcg agtgtgaagg gtatgatttc tagtggtgaa cacctttagt tacgtaatgt
     8821 tttcattgct gttttacttg agatttcgat tgagaaaaag gtatttaata gctcgaatca
     8881 atgtgagaac agagagaaga tgttcttccc taactcgaaa ggtatatgag gcttgtgttt
     8941 cttaggagaa ttattattct tttgttatgt tgcgcttgta gttggaaaag gtgaagagac
     9001 aaaagctgga attgtgagcg gataacaagc tcaacacttg aaatttagga aagagcagaa
     9061 tttggcaaaa aaaataaaaa aaaaataaac acacatactc atcgag
//
