LOCUS       sequence-8430-ex        6367 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 6367)
  AUTHORS   Besmer E, Market E, Papavasiliou FN
  TITLE     The transcription elongation complex directs activation-induced 
            cytidine deaminase-mediated DNA deamination.
  JOURNAL   Mol Cell Biol. 2006 Jun . 26(11):4378-85.
  PUBMED    16705187
REFERENCE   2  (bases 1 to 6367)
  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: "Mol Cell 
            Biol. 2006 Jun . 26(11):4378-85."
COMMENT     Sequence Label: ptac::ccdB
FEATURES             Location/Qualifiers
     source          1..6367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..158
                     /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             159..1241
                     /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     complement(178..197)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1254..1275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1290..1295,1296..1313,1314..1320)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1328..1344
                     /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     1352..1368
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1352..1368
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1364..1891
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1384..1401)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1384..1400)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1393..1415)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1993..1998,1999..2014,2015..2021)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2029..2045
                     /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)."
     CDS             2077..2379
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2258..2277
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2633..3424)
                     /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"
     CDS             complement(4666..5616)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5664..5886
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gaattcgagc tcggtaccag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
       61 tttcacaccg cataaattcc gacaccatcg aatggtgcaa aacctttcgc ggtatggcat
      121 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
      181 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
      241 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
      301 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
      361 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
      421 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
      481 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
      541 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
      601 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
      661 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
      721 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
      781 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
      841 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
      901 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
      961 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgtcaacca
     1021 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     1081 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     1141 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     1201 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     1261 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     1321 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga
     1381 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     1441 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1501 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     1561 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     1621 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtaaccta
     1681 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     1741 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
     1801 cgttggaatt agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
     1861 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
     1921 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
     1981 tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat
     2041 aacaatttca cacaggaaac agtattcatg ggatcccagt ttaaggttta cacctataaa
     2101 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgccgggg
     2161 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     2221 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     2281 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     2341 atcaaaaacg ccattaacct gatgttctgg ggaatataaa agcttagtaa agccctcgct
     2401 agattttaat gcggatgttg cgattacttc gccaactatt gcgataacaa gaaaaagcca
     2461 gcctttcatg atatatctcc caatttgtgt agggcttatt atgcacgctt aaaaataata
     2521 aaagcagact tgacctgata gtttggctgt gagcaattat gtgcttagtg catctaacgc
     2581 ttgagttaag ccgcgccgcg aagcggcgtc ggcttgaacg aattgttaga cattatttgc
     2641 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
     2701 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     2761 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     2821 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     2881 cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     2941 gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc
     3001 tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga
     3061 agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg
     3121 gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa
     3181 tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg
     3241 ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca
     3301 ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc
     3361 gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc
     3421 tcatgatgtt taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc
     3481 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     3541 actgtacccc aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     3601 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     3661 gcttacgaac cgaacaggct tatgtccact gggttcgtgc cttcatccgt ttccacggtg
     3721 tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga
     3781 acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct
     3841 acggcaaggt gctgtgcacg gatctgccct ggcttcagga gatcggaaga cctcggccgt
     3901 cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg
     3961 aaggcgagca tcgtttgttc gcccagcttc tgtatggaac gggcatgcgg atcagtgagg
     4021 gtttgcaact gcgggtcaag gatctggatt tcgatcacgg cacgatcatc gtgcgggagg
     4081 gcaagggctc caaggatcgg gccttgatgt tacccgagag cttggcaccc agcctgcgcg
     4141 agcaggggaa ttacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt
     4201 tatcagaatc gcagatccgg cttcaggttt gccggctgaa agcgctattt cttccagaat
     4261 tgccatgatt ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctttgat
     4321 tcgataagca gcatcgcctg tttcaggctg tctatgtgtg actgttgagc tgtaacaagt
     4381 tgtctcaggt gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta
     4441 ttaggtgtta catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagcttta
     4501 aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct
     4561 gtgcatatgg acagttttcc ctttgatatc taacggtgaa cagttgttct acttttgttt
     4621 gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga tccttccgta
     4681 tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc
     4741 attgagatca tgcttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg
     4801 aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc
     4861 tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc
     4921 ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg
     4981 gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg
     5041 tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat
     5101 gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag
     5161 ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg
     5221 ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact
     5281 aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc
     5341 aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc
     5401 tttagctaat acaccataag cattttccct actgatgttc atcatctgag cgtattggtt
     5461 ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag
     5521 tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc
     5581 tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt
     5641 ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg
     5701 agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag
     5761 accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat
     5821 aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta
     5881 taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc
     5941 tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg
     6001 caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc
     6061 gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca
     6121 ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg
     6181 gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc
     6241 agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca
     6301 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta
     6361 ctgtcgg
//
