LOCUS       sequence-345134-        5260 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes sgRNA target 2300 and Tetracycline resistance transposon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5260)
  AUTHORS   Trujillo Rodriguez L, Ellington AJ, Reisch CR, Chevrette MG
  TITLE     CRISPR-Associated Transposase for Targeted Mutagenesis in Diverse 
            Proteobacteria.
  JOURNAL   ACS Synth Biol. 2023 Jun 27. doi: 10.1021/acssynbio.3c00065.
  PUBMED    37368499
REFERENCE   2  (bases 1 to 5260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2023 Jun 27. doi: 10.1021/acssynbio.3c00065."
COMMENT     Sequence Label: pUCSh3-2300
FEATURES             Location/Qualifiers
     source          1..5260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(231..265)
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     terminator      complement(279..350)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        395..499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(500..568,569..1360)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(718..737)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1531..2119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2020..2039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2273..2290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2655..2749
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2852..2938
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2986..3033
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(3301..4500)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     protein_bind    4728..4775
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(5209..5236)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ctttcaaccc atttagggtt cccccttgat cgggggtgat tcagtagcca ctacagcagt
       61 acatccgcgc ccttattgct gggagcgcac ctatcctgtc tattgacagc agcatcaggg
      121 tgagcagcta ccagggtcag aaagcaagac tgtatttata cagtcaaact aaccctcatt
      181 taacaaaatt cagaggctgc aagcagcatg aattgcggcg ctattaatat gctagcatta
      241 tacctaggac tgagctagct gtcagggtct ctctagtaga gagcgttcac cgacaaacaa
      301 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggag
      361 atccttacca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      421 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      481 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      541 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      601 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      661 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      721 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      781 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      841 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      901 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      961 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1021 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1081 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1141 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1201 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1261 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1321 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1381 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1441 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1501 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1561 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2281 agtgagcgag gaagcggaca gggaagagcg caccggctgg taacaggatt agcagagcga
     2341 ggtatgtagg cggtgctaca agattgtact gagagtgcac catacaaaac cattgtagag
     2401 aggtagcatt gctacgtctc tacgtgtaca gtgactaatt atatgtcgtt gtgacaaatt
     2461 attgtcatca gtaaaatcct tatacagtat agattatagc gctttggcag ttttagcata
     2521 acctctttgc agtgacaaaa tagatgtcgt tgtccgtgat tgtgacaaat tagctgtcgc
     2581 tttgcaagat aggaaaaagc ttttgtgtat tttcataatg acaaattgac tgtcgcctta
     2641 attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     2701 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagctagac
     2761 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     2821 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2881 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2941 gccgctagga gcttgcggcc cggacgatct cgaattgggg atcttgaagt acctattccg
     3001 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaagct gatgtgctcc
     3061 gggggatctc gcgactcgaa cctacagttg tttccttact gggctttctc agcccgggga
     3121 ccgccgtgtt gctaggatgg ttgttcttgg atcagacgct gagtgcgctt caaatcatcg
     3181 gcgtcctgct cgtgatcggg agtatctggc tgggccaacg ttccaaccgc actcctaggg
     3241 cgcgtatagc ttgccggaag tcgccttgac ccgcatggca taggcctatc gtttccacga
     3301 tcagcgatcg gctcgttgcc ctgcgccgct ccaaagcccg cgacgcagcg ccggcaggca
     3361 gagcaagtag agggcagcgc ctgcaatcca tgcccacccg ttccacgttg ttatagaagc
     3421 cgcatagatc gccgtgaaga ggaggggtcc gacgatcgag gtcaggctgg tgagcgccgc
     3481 cagtgagcct tgcagctgcc cctgacgttc ctcatccacc tgcctggaca acattgcttg
     3541 cagcgccggc attccgatgc cacccgaagc aagcaggacc atgatcggga acgccatcca
     3601 tccccgtgtc gcgaaggcaa gcaggatgta gcctgtgccg tcggcaatca ttccgagcat
     3661 gagtgcccgc ctttcgccga gccgggcggc tacagggccg gtgatcattg cctgggcgag
     3721 tgaatgcaga atgccaaatg cggcaagcga aatgccgatc gtggtcgcgt cccagtgaaa
     3781 gcgatcctcg ccgaaaatga cccaaagcgc ggccggcacc tgtccgacaa gttgcatgat
     3841 gaagaagacc gccatcaggg cggcgacgac ggtcatgccc cgggcccacc ggaacgaagc
     3901 gagcgggttg agagcctccc ggcgtaacgg ccggcgttcg cctttgtgcg actccggcaa
     3961 aaggaaacag cccgtcagga aattgaggcc gttcaaggct gccgcggcga agaacggagc
     4021 gtggggggag aaaccgccca tcagcccacc gagcacaggt cccgcgacca tcccgaaccc
     4081 gaaacaggcg ctcatgaagc cgaagtgccg cgcgcgctca tcgccatcag tgatatcggc
     4141 aatataagcg ccggctaccg ccccagtcgc cccggtgatg ccggccacga tccgcccgat
     4201 atagagaacc caaaggaaag gcgctgtcgc catgatggcg tagtcgacag tggcgccggc
     4261 cagcgagacg agcaagattg gccgccgccc gaaacgatcc gacagcgcgc ccagcacagg
     4321 tgcgcaggca aattgcacca acgcatacag cgccagcaga atgccatagt gggcggtgac
     4381 gtcgttcgag tgaaccagat cgcgcaggag gcccggcagc accggcataa tcaggccgat
     4441 gccgacagcg tcgagcgcga cagtgctcag aattacgatc aggggtatgt tgggtttcac
     4501 gtctggcctc cggaccagcc tccgctggtc cgattgaacg cgcggattct ttatcactga
     4561 taagttggtg gacgtccaga tagcccagta gctgacattc atccggggtc tgcaccgttt
     4621 ctgcggactg gctttctacg tgttccgctt cctttagcag cccttgcgcc atgcatttgt
     4681 taaatcgaat tatccattgc tgttgacaaa gggaatcagg ggatcttgaa gttcctattc
     4741 cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag ctaattcgag
     4801 atcatgcatg agctcactag tggatccccc tcagtaacta ttaaacttag gggtgggttg
     4861 aaagcaagtc cttttatccg cttgttttaa ttgctttgta taataattgc agagcatatt
     4921 atattgatga catttaattt gtcatcaatt aattaagcaa cgctgatggg tcacgacgac
     4981 aattaaatag tcacaatgac attaatctgt caccgacgac agataatttg tcactgtaca
     5041 ctacgccttt tgtggagatg tctaataaac taaaaatttt ttactttccg ctgcataacc
     5101 ctgcttcggg gtcattgacg aaagggcctc gtgatctcag gccgcctagg ccggtttccc
     5161 gactggaaag cggggttccg cgcacatttc cccgaaaagt gccacctgcg cagaaaggcc
     5221 cacccgaagg tgagccctgc agtgcttcgc ccgctacccc
//
