LOCUS       sequence-367503-        5706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses retron Eco1 RT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5706)
  AUTHORS   Bhattarai-Kline S, Lear SK, Fishman CB, Lopez SC, Lockshin ER, 
            Schubert MG, Nivala J, Church GM, Shipman SL
  TITLE     Recording gene expression order in DNA by CRISPR addition of retron 
            barcodes.
  JOURNAL   Nature. 2022 Aug;608(7921):217-225. doi: 10.1038/s41586-022-04994-6.
            Epub 2022 Jul 27.
  PUBMED    35896746
REFERENCE   2  (bases 1 to 5706)
  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"; doi: 
            "10.1038/s41586-022-04994-6"; journalName: "Nature"; date: 
            "2022-08"; volume: "608"; issue: "7921"; pages: "217-225"
COMMENT     Sequence Label: pSLS.402
FEATURES             Location/Qualifiers
     source          1..5706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(118..135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(252..797)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(332..351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1035..1826)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVITEVSTQLSEVVGAIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(2221..2238)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2391..2477
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             3885..3893
                     /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(4500..4594)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(5367..5471)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5539..5557
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      5625..5672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagacgg atcgcttcat gtggtaggag
       61 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
      121 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
      181 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
      241 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
      301 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
      361 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
      421 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
      481 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
      541 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
      601 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
      661 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
      721 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
      781 gaccaaaacg atctcaagaa gatcatctta ttaaggggtc tgacgctcag tggaacgaaa
      841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccagc
      901 ggtatcatca acaggcttac ccgtcttact gtcaagagga catccggtca aataaaacga
      961 aaggctcagt cgaaagactg ggcctttcgt tttgctgagg agacttaggg acccttcgaa
     1021 ccccagagtc ccgcttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     1081 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     1141 tgtctagctt caagtatgac gggctgatat tgggccggca ggcgctccat tgcccagtcg
     1201 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     1261 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     1321 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     1381 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     1441 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     1501 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     1561 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     1621 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aactagcaaa
     1681 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     1741 agcaacgtcg gttcgagatg gcgctcgatg gcgccaacta cctctgatag ttgagtcgat
     1801 acttcggtga tcaccgcttc cctcataatg tttaactttg ttttagggcg actgccctgc
     1861 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccatggcgta acgcgcttgc
     1921 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcatagca agccatgaaa
     1981 accgccactg caccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     2041 cgcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     2101 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     2161 ggcgccccag ctggcaattc cgacgtctgg ctgttttggc ggatgagaga agattttcag
     2221 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     2281 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     2341 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     2401 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2461 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     2521 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     2581 gtcaggatgg cctttttgcg tttctacaaa ctggattgaa tataaccgac gtgactgtta
     2641 catttaggtg gctaaacccg tcaacattta actttaagaa ggagatatac atatgaaatc
     2701 cgctgaatat ttgaacactt ttagattgag aaatctcggc ctacctgtca tgaacaattt
     2761 gcatgacatg tctaaggcga ctcgcatatc tgttgaaaca cttcggttgt taatctatac
     2821 agctgatttt cgctatagga tctacactgt agaaaagaaa ggcccagaga agagaatgag
     2881 aaccatttac caaccttctc gagaacttaa agccttacaa ggatgggttc tacgtaacat
     2941 tttagataaa ctgtcgtcat ctcctttttc tattggattt gaaaagcacc aatctatttt
     3001 gaataatgct accccgcata ttggggcaaa ctttatactg aatattgatt tggaggattt
     3061 tttcccaagt ttaactgcta acaaagtttt tggagtgttc cattctcttg gttataatcg
     3121 actaatatct tcagttttga caaaaatatg ttgttataaa aatctgctac cacaaggtgc
     3181 tccatcatca cctaaattag ctaatctaat atgttctaaa cttgattatc gtattcaggg
     3241 ttatgcaggt agtcggggct tgatatatac gagatatgcc gatgatctca ccttatctgc
     3301 acagtctatg aaaaaggttg ttaaagcacg tgatttttta ttttctataa tcccaagtga
     3361 aggattggtt attaactcaa aaaaaacttg tattagtggg cctcgtagtc agaggaaagt
     3421 tacaggttta gttatttcac aagagaaagt tgggataggt agagaaaaat ataaagaaat
     3481 tagagcaaag atacatcata tattttgcgg taagtcttct gagatagaac acgttagggg
     3541 atggttgtca tttattttaa gtgtggattc aaaaagccat aggagattaa taacttatat
     3601 tagcaaatta gaaaaaaaat atggaaagaa ccctttaaat aaagcgaaga cctaattaat
     3661 taacggcact cctcagcaaa tataatgacc ctcttgataa cccaagaggg cattttttaa
     3721 tgcccatggc gtttaccaca gctaacacca cgtcgtccct atctgctgcc ctaggtctat
     3781 gagtggttgc tggataactt tacgggcatg cataaggctc gtaggctata ttcagggaga
     3841 ccacaacggt ttccctctac aaataatttt gtttaacttt gaaataagga ggtaatacaa
     3901 atgccgcgtc cgaaactgaa atctgacgac gaagttctgg aagcggcgac cgttgttctg
     3961 aaacgttgcg gtccgatcga attcaccctg tctggtgttg cgaaagaagt tggtctgtct
     4021 cgtgcggcgc tgatccagcg tttcaccaac cgtgacaccc tgctggttcg tatgatggaa
     4081 cgtggtgttg aacaggttcg tcactacctg aacgcgatcc cgatcggtgc gggtccgcag
     4141 ggtctgtggg aattcctgca ggttctggtt cgttctatga acacccgtaa cgacttctct
     4201 gttaactacc tgatctcttg gtacgaactg caggttccgg aactgcgtac cctggcgatc
     4261 cagcgtaacc gtgcggttgt tgaaggtatc cgtaaacgtc tgccgccggg tgcgccggcg
     4321 gcggcggaac tgctgctgca ctctgttatc gcgggtgcga ccatgcagtg ggcggttgac
     4381 ccggacggtg aactggcgga ccacgttctg gcgcagatcg cggcgatcct gtgcctgatg
     4441 ttcccggaac acgacgactt ccagctgctg cagccgcacg cgtaataact agtgcttgga
     4501 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     4561 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgtaaa cttggtctga
     4621 cagcccttaa cttacttatt aaataattta tagctattga aaagagataa gaattgttca
     4681 aagctaatat tgtttaaatc gtcaattcct gcatgtttta aggaattgtt aaattgattt
     4741 tttgtaaata ttttcttgta ttctttgtta acccatttca taacgaaata attatacttt
     4801 tgtttatctt tgtgtgatat tcttgatttt tttctactta atctgataag tgagctattc
     4861 actttaggtt taggatgaaa atattctctt ggaaccatac ttaatataga aatatcaact
     4921 tctgccatta aaagtaatgc caatgagcgt tttgtattta ataatctttt agcaaacccg
     4981 tattccacga ttaaataaat ctcattagct atactatcaa aaacaatttt gcgtattata
     5041 tccgtactta tgttataagg tatattacca tatattttat aggattggtt tttaggaaat
     5101 ttaaactgca atatatcctt gtttaaaact tggaaattat cgtgatcaac aagtttattt
     5161 tctgtagttt tgcataattt atggtctatt tcaatggcag ttacgaaatt acacctcttt
     5221 actaattcaa gggtaaaatg gccttttcct gagccgattt caaagatatt atcatgttca
     5281 tttaatctta tatttgtcat tattttatct atattatgtt ttgaagtaat aaagttttga
     5341 ctgtgtttta tatttttctc gttcatactc ttcctttttc aatattattg aagcatttat
     5401 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5461 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5521 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca ctaagcgact
     5581 aaaaaattga atgtaggaaa ccaacatgcc agttctagca ataactagca taaccccttg
     5641 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcgaga agcacacggt
     5701 cacact
//
