LOCUS       sequence-8434-ex        6861 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 6861)
  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 6861)
  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-KanL94P
FEATURES             Location/Qualifiers
     source          1..6861
                     /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             2078..2872
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2132..2151)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2742..2761
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(3127..3918)
                     /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(5160..6110)
                     /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      6158..6380
                     /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 ggatccgatg attgaacaag atggattgca
     2101 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2161 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2221 tgtcaagacc gacctgtccg gtgccctgaa tgaactccaa gacgaggcag cgcggctatc
     2281 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     2341 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     2401 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     2461 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     2521 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     2581 cgaactgttc gccaggctca aggcgcggat gcccgacggc gaggatctcg tcgtgaccca
     2641 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     2701 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     2761 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     2821 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaaaagctta
     2881 gtaaagccct cgctagattt taatgcggat gttgcgatta cttcgccaac tattgcgata
     2941 acaagaaaaa gccagccttt catgatatat ctcccaattt gtgtagggct tattatgcac
     3001 gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa ttatgtgctt
     3061 agtgcatcta acgcttgagt taagccgcgc cgcgaagcgg cgtcggcttg aacgaattgt
     3121 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     3181 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     3241 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     3301 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     3361 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     3421 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     3481 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     3541 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     3601 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     3661 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     3721 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     3781 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     3841 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     3901 gatcaccgct tccctcatga tgtttaactt tgttttaggg cgactgccct gctgcgtaac
     3961 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     4021 tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga aaaccgccac
     4081 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     4141 ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc gtgccttcat
     4201 ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct
     4261 gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc
     4321 cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc aggagatcgg
     4381 aagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat
     4441 cctcggtttt ctggaaggcg agcatcgttt gttcgcccag cttctgtatg gaacgggcat
     4501 gcggatcagt gagggtttgc aactgcgggt caaggatctg gatttcgatc acggcacgat
     4561 catcgtgcgg gagggcaagg gctccaagga tcgggccttg atgttacccg agagcttggc
     4621 acccagcctg cgcgagcagg ggaattacgg gttttgctgc ccgcaaacgg gctgttctgg
     4681 tgttgctagt ttgttatcag aatcgcagat ccggcttcag gtttgccggc tgaaagcgct
     4741 atttcttcca gaattgccat gattttttcc ccacgggagg cgtcactggc tcccgtgttg
     4801 tcggcagctt tgattcgata agcagcatcg cctgtttcag gctgtctatg tgtgactgtt
     4861 gagctgtaac aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg
     4921 tttcacctgt tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat
     4981 ggtgaacagc tttaaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta
     5041 caccgttttc atctgtgcat atggacagtt ttccctttga tatctaacgg tgaacagttg
     5101 ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
     5161 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
     5221 catgagaacg aaccattgag atcatgctta ctttgcatgt cactcaaaaa ttttgcctca
     5281 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
     5341 cgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
     5401 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
     5461 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
     5521 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
     5581 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
     5641 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
     5701 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
     5761 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
     5821 cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc
     5881 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
     5941 tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc
     6001 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
     6061 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
     6121 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
     6181 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     6241 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     6301 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     6361 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     6421 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     6481 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
     6541 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
     6601 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     6661 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
     6721 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     6781 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     6841 gcttacccgt cttactgtcg g
//
