LOCUS       sequence-429180-        9908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminus of ABE, and untargeted Bacterial Mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 6004 to 6038)
  AUTHORS   Qi T, Wang Y, Yang Y, Gao S, Liu J, Huang Q, Tian Y, Tang J, Zheng 
            WV, Wang Y
  TITLE     Phage-assisted evolution of compact Cas9 variants targeting a simple
            NNG PAM.
  JOURNAL   Nat Chem Biol. 2023 Dec 5. doi: 10.1038/s41589-023-01481-5.
  PUBMED    38052959
REFERENCE   2  (bases 1 to 9908)
  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: "Nat Chem 
            Biol. 2023 Dec 5. doi: 10.1038/s41589-023-01481-5."
COMMENT     Sequence Label: MP6-SlugABEN-738
FEATURES             Location/Qualifiers
     source          1..9908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        286..570
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     493..512
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3378..3632
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3662..4288
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     terminator      complement(4315..4346)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4362..5021)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      complement(5174..5912)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     promoter        6004..6038
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             6046..6054
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      complement(9206..9237)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(9289..9908,1..259))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
       61 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
      121 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
      181 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
      241 cattttgcgc ttcagccata cttttcatac tcccaccatt cagagaagaa accaattgtc
      301 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaacccaac
      361 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa
      421 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg
      481 tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct
      541 ttttatcgca actctctact gtttctccat acccgttttt ttggacgcgt acaactcaag
      601 tctgacataa atgaccgcta tgagcactgc aattacacgc cagatcgttc tcgctaccgc
      661 aaccaccggt atgaaccaga ttggtgcgca ctatgaaggc cacaagatca ttgagattgg
      721 tgccgttgaa gtggtgaacc gtcgcctgac gggcaataac ttccatgttt atctcaaacc
      781 cgatcggctg gtggatccgg aagcctttgg cgtacatggt attgccgatg aatttttgct
      841 cgataagccc acgtttgccg aagtagccga tgagttcatg gactatattc gcggcgcgga
      901 gttggtgatc cataacgcag cgttcgatat cggctttatg gactacgagt tttcgttgct
      961 taagcgcgat attccgaaga ccaatacttt ctgtaaggtc accgatagcc ttgcggtggc
     1021 gaggaaaatg tttcccggta agcgcaacag cctcgatgcg ttatgtgctc gctacgaaat
     1081 agataacagt aaacgaacgc tgcacggggc attactcgat gcccagatcc ttgcggaagt
     1141 ttatctggcg atgaccggtg gtcaaacgtc gatggctttt gcgatggaag gagagacaca
     1201 acagcaacaa ggtgaagcaa caattcagcg cattgtacgt caggcaagta agttacgcgt
     1261 tgtttttgcg acagatgaag agattgcagc tcatgaagcc cgtctcgatc tggtgcagaa
     1321 gaaaggcgga agttgcctct ggcgagcata atttaatatc agtaaaccgg acataaccca
     1381 tgaagaaaaa tcgcgctttt ttgaagtggg cagggggcaa gtatcccctg cttgatgata
     1441 ttaaacggca tttgcccaag ggcgaatgtc tggttgagcc ttttgtaggt gccgggtcgg
     1501 tgtttctcaa caccgacttt tctcgttata tccttgccga tatcaatagc gacctgatca
     1561 gtctctataa cattgtgaag atgcgtactg atgagtacgt acaggccgca cgcgagctgt
     1621 ttgttcccga aacaaattgc gccgaggttt actatcagtt ccgcgaagag ttcaacaaaa
     1681 gccaggatcc gttccgtcgg gcggtactgt ttttatattt gaaccgctac ggttacaacg
     1741 gcctgtgtcg ttacaatctg cgcggtgagt ttaacgtgcc gttcggccgc tacaaaaaac
     1801 cctatttccc ggaagcagag ttgtatcact tcgctgaaaa agcgcagaat gcctttttct
     1861 attgtgagtc ttacgccgat agcatggcgc gcgcagatga tgcatccgtc gtctattgcg
     1921 atccgcctta tgcaccgctg tctgcgaccg ccaactttac ggcgtatcac acaaacagtt
     1981 ttacgcttga acaacaagcg catctggcgg agatcgccga aggtctggtt gagcgccata
     2041 ttccagtgct gatctccaat cacgatacga tgttaacgcg tgagtggtat cagcgcgcaa
     2101 aattgcatgt cgtcaaagtt cgacgcagta taagcagcaa cggcggcaca cgtaaaaagg
     2161 tggacgaact gctggctttg tacaaaccag gagtcgtttc acccgcgaaa aaataattca
     2221 gctaagacac tgcactggat taagatgaaa acgattgaag ttgatgatga actctacagc
     2281 tatattgcca gccacactaa gcatatcggc gagagcgcat ccgacatttt acggcgtatg
     2341 ttgaaatttt ccgccgcatc acagcctgct gctccggtga cgaaagaggt tcgcgttgcg
     2401 tcacctgcta tcgtcgaagc gaagccggtc aaaacgatta aagacaaggt tcgcgcaatg
     2461 cgtgaacttc tgctttcgga tgaatacgca gagcaaaagc gagcggtcaa tcgctttatg
     2521 ctgctgttgt ctacactata ttctcttgac gcccaggcgt ttgccgaagc aacggaatcg
     2581 ttgcacggtc gtacacgcgt ttactttgcg gcagatgaac aaacgctgct gaaaaatggt
     2641 aatcagacca agccgaaaca tgtgccaggc acgccgtatt gggtgatcac caacaccaac
     2701 accggccgta aatgcagcat gatcgaacac atcatgcagt cgatgcaatt cccggcggaa
     2761 ttgattgaga aggtttgcgg aactatctaa cggctgaaat taatgaggtc atacccaaat
     2821 ggatagttcg tttacgccca ttgaacaaat gctaaaattt cgcgccagcc gccacgaaga
     2881 ttttccttat caggagatcc ttctgactcg tctttgcatg cacatgcaaa gcaagctgct
     2941 ggagaaccgc aataaaatgc tgaaggctca gggaattaac gagacgttgt ttatggcgtt
     3001 gattacgctg gagtctcagg aaaaccacag tattcagcct tctgaattaa gttgtgctct
     3061 tggatcatcc cgtaccaacg cgacgcgtat tgccgatgaa ctggaaaaac gcggttggat
     3121 cgaacgtcgt gaaagcgata acgatcgccg ctgcctgcat ctgcaattaa cggaaaaagg
     3181 tcacgagttt ttgcgcgagg ttttaccacc gcagcataac tgcctgcatc aactctggtc
     3241 cgcgctcagc acaacagaaa aagatcagct cgagcaaatc acccgcaaat tgctctcccg
     3301 tctcgaccag atggaacaag acggtgtggt tctcgaagcg atgagctaat aatacaaaaa
     3361 ttaggaggaa tttcaacatg acaaatttat ctgacatcat tgaaaaagaa acaggaaaac
     3421 aactagtgat tcaagaatca attctaatgt taccagaaga agtagaggaa gtaattggga
     3481 ataaaccaga aagtgatatt ttagttcata ctgcttatga tgaaagtaca gatgaaaatg
     3541 taatgctatt aacttcagat gctccagaat ataaaccttg ggctttagta attcaagaca
     3601 gtaatggaga aaataaaatt aaaatgttat aagtcgagat taagtaaacc ggaatctgaa
     3661 gatgaccgac gcggaatacg ttcgtatcca cgaaaaactg gacatctaca ccttcaaaaa
     3721 acagttcttc aacaacaaaa aatctgtttc tcaccgttgc tacgttctgt tcgaactgaa
     3781 acgtcgtggt gaacgtcgtg cgtgcttctg gggttacgcg gttaacaaac cgcagtctgg
     3841 taccgaacgt ggtatccacg cggaaatctt ctctatccgt aaagttgaag aatacctgcg
     3901 tgacaacccg ggtcagttca ccatcaactg gtactcttct tggtctccgt gcgcggactg
     3961 cgcggaaaaa atcctggaat ggtacaacca ggaactgcgt ggtaacggtc acaccctgaa
     4021 aatctgggcg tgcaaactgt actacgaaaa aaacgcgcgt aaccagatcg gtctgtggaa
     4081 cctgcgtgac aacggtgttg gtctgaacgt tatggtttct gaacactacc agtgctgccg
     4141 taaaatcttc atccagtctt ctcacaacca gctgaacgaa aaccgttggc tggaaaaaac
     4201 cctgaaacgt gcggaaaaac gtcgttctga actgtctatc atgatccagg ttaaaatcct
     4261 gcacaccacc aaatctccgg cggtttaaac ttaattaacg gcactcctca gccaagtcaa
     4321 aagcctccgg tcggaggctt ttgactacat gcccatggcg tttacgcccc gccctgccac
     4381 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     4441 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     4501 cccatagtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     4561 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     4621 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     4681 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     4741 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     4801 cggaactccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     4861 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     4921 ttataggtac attgagtaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     4981 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     5041 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     5101 ttggaacctc ttacgtgcca agccaaatag gccgtcactc ggtcgctacg ctccgggcgt
     5161 gagactgcgg cgggcgctgc ggacacatac aaagttaccc acagattccg tggataagca
     5221 ggggactaac atgtgaggca aaacagcagg gccgcgccgg tggcgttttt ccataggctc
     5281 cgccctcctg ccagagttca cataaacaga cgcttttccg gtgcatctgt gggagccgtg
     5341 aggctcaacc atgaatctga cagtacgggc gaaacccgac aggacttaaa gatccccacc
     5401 gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg accctgccgt ttaccggata
     5461 cctgttccgc ctttctccct tacgggaagt gtggcgcttt ctcatagctc acacactggt
     5521 atctcggctc ggtgtaggtc gttcgctcca agctgggctg taagcaagaa ctccccgttc
     5581 agcccgactg ctgcgcctta tccggtaact gttcacttga gtccaacccg gaaaagcacg
     5641 gtaaaacgcc actggcagca gccattggta actgggagtt cgcagaggat ttgtttagct
     5701 aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc tacactggaa ggacagattt
     5761 ggttgctgtg ctctgcgaaa gccagttacc acggttaagc agttccccaa ctgacttaac
     5821 cttcgatcaa accacctccc caggtggttt tttcgtttac agggcaaaag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac tgaaccgctc tagatttcag
     5941 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc aggaggaaga ggacatccgg
     6001 tgtttgacgg ctagctcagt cctaggtaca gtgctagccg agctctaagg aggttataaa
     6061 aaatgtctga ggtggagttt tcccacgagt actggatgag acatgccctg accctggcca
     6121 agagggcacg ggatgagagg gaggtgcctg tgggagccgt gctggtgctg aacaatagag
     6181 tgatcggcga gggctggaac agagccatcg gcctgcacga cccaacagcc catgccgaaa
     6241 ttatggccct gagacagggc ggcctggtca tgcagaacta cagactgatt gacgccaccc
     6301 tgtacgtgac attcgagcct tgcgtgatgt gcgccggcgc catgatccac tctaggatcg
     6361 gccgcgtggt gtttggcgtg aggaactcaa aaagaggcgc cgcaggctcc ctgatgaacg
     6421 tgctgaacta ccccggcatg aatcaccgcg tcgaaattac cgagggaatc ctggcagatg
     6481 aatgtgccgc cctgctgtgc gatttctatc ggatgcctag acaggtgttc aatgctcaga
     6541 agaaggccca gagctccatc aactccggag gatctagcgg aggctcctct ggctctgaga
     6601 cacctggcac aagcgagagc gcaacacctg aaagcagcgg gggcagcagc ggggggtcaa
     6661 accaaaaatt catactggga ctggccatcg gaatcaccag cgtgggctac ggcctgatcg
     6721 actacgagac aaagaatatc atcgatgccg gcgttagact gttccccgag gccaacgtgg
     6781 aaaacaacga gggaagaagg tccaaacgtg gaagcagaag actgaagcga cgccgcattc
     6841 acagacttga acgggtgaag aagctgctcg aggattataa tctgctggat cagtcccaga
     6901 ttcctcagtc tacaaacccc tacgccatcc gcgtgaaggg cctgtctgaa gccctgagca
     6961 aggacgaact cgtgattgcc ctgctccata tcgccaagag aagaggcatc cacaagatcg
     7021 acgtgatcga cagcaacgac gacgtgggga acgagctcag caccaaggaa cagctgaata
     7081 agaacagcaa gctgctgaaa gacaaatttg tgtgccagat ccagctggaa agaatgaatg
     7141 agggccaggt gcggggagag aaaaaccggt tcaagaccgc tgatatcatc aaggaaatca
     7201 tccagctgct gaatgtgcag aagaacttcc accagctgga cgagaacttc atcaacaagt
     7261 acatcgaact ggttgagatg agacgggaat acttcgaggg ccccggcaag ggcagtccat
     7321 atggctggga aggcgaccct aaggcttggt acgagacact gatgggccac tgcacctact
     7381 tcccagatga gctgagaagc gtgaaatacg cctacagtgc cgacctgttc aacgctctga
     7441 acgacctgaa caacctggtc atccaaagag atggactgtc taagctcgag tatcatgaga
     7501 agtatcacat catcgagaac gtgttcaagc agaagaagaa acctacactg aagcagatcg
     7561 ccaatgagat caatgtcaac cctgaagata tcaagggcta cagaatcaca aagtctggca
     7621 agccccagtt taccgagttt aagctctacc acgacctgaa aagcgtgctg tttgaccaga
     7681 gcatcctgga gaacgaagac gtgctggacc agatcgctga gatcctgacc atctaccagg
     7741 acaaggatag catcaaatct aagctgacgg aactggacat cctgctgaac gaggaagata
     7801 aggaaaacat cgcccagctg actggctaca ccgggaccca ccggctcagc ctgaaatgca
     7861 tccggctggt cctggaagag cagtggtatt ctagccggaa ccagatggaa atcttcaccc
     7921 acctgaacat taagcctaag aagatcaacc tgacagccgc caacaagatc ccgaaggcta
     7981 tgatcgacga gttcatcctg agccctgtgg tgaagaggac cttcggccag gccattaacc
     8041 ttattaacaa gatcatagaa aagtacggcg tgcctgaaga tatcatcatc gagctggcca
     8101 gagaaaataa tagcaaggac aagcagaagt tcatcaatga gatgcagaaa aagaacgaga
     8161 acaccagaaa gagaattaac gaaatcatcg gcaagtatgg caaccagaac gccaagagac
     8221 tggtcgagaa gattagactg cacgacgagc aggagggcaa gtgcctgtac tcactggaaa
     8281 gcatccctct ggaggacctg ctgaacaacc ccaaccacta cgaggtggac cacatcattc
     8341 caagatctgt gtccttcgac aactcttacc acaacaaagt gctcgtgaag cagagcgagg
     8401 cctccaaaaa atccaacctg accccttacc agtactttaa cagcggcaag tccaagctct
     8461 cttacaacca gtttaaacaa cacatcctga acctgagcaa gtcccaggat agaatcagca
     8521 aaaaaaagaa agagtatctg ctggaagaac gggacatcaa caagttcgag gtgcaaaaag
     8581 agttcatcaa tagaaacctg gtggataccc ggtacgccac aagagagctg acaaactacc
     8641 tgaaggccta cttcagcgcc aacaatatga acgtgaaggt gaaaacgatc aacggcagct
     8701 tcaccgatta cctgcggaaa gtgtggaagt ttaagaagga acggaaccac ggctacaagc
     8761 accacgccga ggacgccctg attatcgcta atgccgattt cctgttcaaa gagaacaaga
     8821 agctgaaagc cgtgaactct gtgctggaaa aacctgagat cgagagcaag tgcctgtcat
     8881 acgaaaccga gatactgaca gtagaatatg gccttctgcc aatcgggaag attgtggaga
     8941 aacggataga atgcacagtt tactctgtcg ataacaatgg taacatttat actcagccag
     9001 ttgcccagtg gcacgaccgg ggagagcagg aagtattcga atactgtctg gaggatggaa
     9061 gtctcattag ggccactaag gaccacaaat ttatgacagt cgatggccag atgctgccta
     9121 tagacgaaat ctttgagcga gagttggacc tcatgcgagt tgacaacctt cctaattaaa
     9181 cttaattaac ggcactcctc agccaagtca aaagcctccg gtcggaggct tttgactaca
     9241 tgcccatggc gtttatttgc cgactacctt ggtggactta gggacccttt atgacaactt
     9301 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     9361 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     9421 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     9481 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     9541 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     9601 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     9661 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     9721 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     9781 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     9841 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     9901 gcgagcct
//
