LOCUS       sequence-51674-e        7993 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 7993)
  AUTHORS   Ivleva NB, Bramlett MR, Lindahl PA, Golden SS
  TITLE     LdpA: a component of the circadian clock senses redox state of the 
            cell.
  JOURNAL   EMBO J. 2005 Mar 23;24(6):1202-10. Epub 2005 Mar 10.
  PUBMED    15775978
REFERENCE   2  (bases 1 to 7993)
  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: "EMBO J."; 
            date: "2005-03-23"; volume: "24(6)"; pages: "1202-10. Epub 2005 Mar 
            10"
COMMENT     Sequence Label: pAM2991
FEATURES             Location/Qualifiers
     source          1..7993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      725..1313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1214..1233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1467..1484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1499..1639
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1565..1584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             3558..4349
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(4814..4841)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4933..5019)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5172..5189
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     misc_feature    5222..5278
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5291..5313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5302..5318
                     /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)."
     promoter        complement(join(5326..5332,5333..5349,5350..5355))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(5587..6669)
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6631..6650
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6670..6747)
                     /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."
ORIGIN
        1 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
       61 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
      241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      421 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      841 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
      901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1441 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1501 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     1561 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     1621 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1741 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag gatcgatccg gcagccggcg
     1801 gagcgctgct ttcttggcaa gcggtcgcca gccccaacgc cagggctgcc agcccgaaac
     1861 agcggggcaa ggcagcttgg aagggcgatc gcagcacggg catggcaatg tctctctgaa
     1921 ggaatgcaga ccttattcgt acagccaggg ttgaatcgtg ggggtccaat cacttagctc
     1981 tgctgggcta aaccagagag caatttcctg ttgtgctgtt tcgattgcat ccgagccatg
     2041 gatgatgttg cggccaatat tgacaccaaa atcaccacgg atggtgcccg gttctgccgt
     2101 cagcggattg gtagcgccga tcaacttgcg agcagccgcc acaacgcctt cgccttccaa
     2161 gacgatcgcc acgatcggcc cagaggtgat gaactcgacg aggccattga agaaggggcg
     2221 ctcgcggtgg acagcatagt gctgttcggc cagctcgcga ctgggcttca gctgctttag
     2281 gcccaccagt ttgaagcctt tttgctcaaa gcggccgatg atcgtaccga ccaaaccccg
     2341 ctgaacgcca tcgggcttga tggcaataaa tgtgcgttcc acagacatct agatagtcct
     2401 caagacgagg caagcattga gcttgccttc ctatggttcg ggatcactgg gattcttgac
     2461 aagcgatcgc ggtcacatcg ctatctctta ggacttcgca gcgggcgagt cggattgacc
     2521 cggtagggat ttcgccagat caatgcccgt ggtttgtttc agcttctcca gcaagctagc
     2581 gatttgggta gcgctgcctt ccccttcgcc aatcacagtg atcgactcca cgtcgatatc
     2641 tggcacggtg cctgaaagcg tgacgagcag ggactcgaag cttgcatgcc tgcaggtcga
     2701 ctctagagct ttatgcttgt aaaccgtttt gtgaaaaaat ttttaaaata aaaaagggga
     2761 cctctagggt ccccaattaa ttagtaatat aatctattaa aggtcattca aaaggtcatc
     2821 caccggatca attcccctgc tcgcgcaggc tgggtgccaa gctctcgggt aacatcaagg
     2881 cccgatcctt ggagcccttg ccctcccgca cgatgatcgt gccgtgatcg aaatccagat
     2941 ccttgacccg cagttgcaaa ccctcactga tccgcatgcc cgttccatac agaagctggg
     3001 cgaacaaacg atgctcgcct tccagaaaac cgaggatgcg aaccacttca tccggggtca
     3061 gcaccaccgg caagcgccgc gacggccgag gtcttccgat ctcctgaagc cagggcagat
     3121 ccgtgcacag caccttgccg tagaagaaca gcaaggccgc caatgcctga cgatgcgtgg
     3181 agaccgaaac cttgcgctcg ttcgccagcc aggacagaaa tgcctcgact tcgctgctgc
     3241 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     3301 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     3361 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     3421 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     3481 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     3541 aaacaaagtt aaacatcatg agggaagcgg tgatcgccga agtatcgact caactatcag
     3601 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     3661 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     3721 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     3781 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     3841 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     3901 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     3961 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     4021 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     4081 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     4141 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     4201 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     4261 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     4321 gcgagatcac caaggtagtc ggcaaataat gtctaacaat tcgttcaagc cgacgccgct
     4381 tcgcggcgcg gcttaactca agcgttagat gcactaagca cataattgct cacagccaaa
     4441 ctatcaggtc aagtctgctt ttattatttt taagcgtgca taataagccc tacacaaatt
     4501 gggagatata tcatgaaagg ctggcttttt cttgttatcg caatagttgg cgaagtaatc
     4561 gcaacatccg cattaaaatc tagcgagggc tttactaagc tgatccggtg gatgaccttt
     4621 tgaatgacct ttaatagatt atattactaa ttaattgggg accctagagg tccccttttt
     4681 tattttaaaa attttttcac aaaacggttt acaagcatac tagaggatcg gcggccgcgg
     4741 atctgggccg ccatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt
     4801 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc
     4861 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc
     4921 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     4981 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     5041 tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     5101 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc
     5161 ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc
     5221 caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattcca
     5281 tggtctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacatta tacgagccgg
     5341 atgattaatt gtcaacagct catttcagaa tatttgccag aaccgttatg atgtcggcgc
     5401 aaaaaacatt atccagaacg ggagtgcgcc ttgagcgaca cgaattatgc agtgatttac
     5461 gacctgcaca gccataccac agcttccgat ggctgcctga cgccagaagc attggtgcac
     5521 cgtgcagtcg atgataagct gtcaaaccag atcaattcgc gctaactcac attaattgcg
     5581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5701 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5761 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttgacggcgg
     5821 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5881 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5941 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     6001 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     6061 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     6121 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     6181 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     6241 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     6301 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     6361 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     6421 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     6481 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     6541 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     6601 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     6661 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     6721 aaaggttttg caccattcga tggtgtcaac gtaaatgcat gccgcttcgc cttcgcgcgc
     6781 gaattgatct gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6841 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6901 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccgcgg cccagatccc
     6961 cgggtaccga gctcgaattt cgagcttctg gagcaggaag atgtcgcggg cattagcacc
     7021 agcggtctgc caagcctccg ccagccgttg ggtcccttcc gcttgagctt ttccatcttc
     7081 gacgatacgg gcggcggccc cccgcgcttc cgcgatcgcc cgtttacaag ctgcctcagc
     7141 tggggcgatc acatcggctt gaagttgctg ctgcacctgt ttgatccgct cctgctgcac
     7201 agggagttct gcttggctac gagcgacttc ggtagcaatg tccgcttcag cttcggccac
     7261 caccgcttcg cgccgcgtca acgcatcctg aatccggcgc tcggcctcgg cttgggcgat
     7321 cgctacatcg cgatcgatcc gacgcagggc cgtgatcttg tcattttcgg ccgtttggat
     7381 cgcagaggca gcctgggcat cggcttcagc aattcgggca tctcgctgca gatcagcccg
     7441 ctgcttgcgt ccactagccg agagataacc gacctcatcg gaaatgttct ggacttgcag
     7501 cgtatcgagg actagaccca gctgctcaag gtcatcctcc gcctcttcca gcagactttt
     7561 ggcaaaggca attttgtcct cgttgatctg ctccggcgtg aggctggcta aaacaccacg
     7621 caagttgcct tcgagggtct ccttggcaat ttgctcgatt tccttacggt ttttgccaag
     7681 cagccgctcg atcgcgttgt ggatggtcgg ttcttcccca gcaatcttga tattggcaac
     7741 gccttcaaca gtcaggggaa tgccgccctt ggagaaggca ttggaaacgc gcaactcaat
     7801 gatcatgttg gtcagatcca tgcggagcgc tttttccagc agaggtaccc gcaggctgct
     7861 gccgcccttg accaagcgat agccaactcg gcggccatca ctactgcggc gactactgcc
     7921 agcaaagatc aaaatttcac tgggttggca gatgtagtag agattgcgca ggactaagct
     7981 gccagccccg gcg
//
