LOCUS       sequence-488066-        2595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA expression vector under tetR transcriptional control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2595)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2595)
  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: 
            "Unpublished"
COMMENT     Sequence Label: ptetgRNA-FRT
FEATURES             Location/Qualifiers
     source          1..2595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        81..156
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             177..206
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             222..239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(295..312)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      465..511
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(564..581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(735..1323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(815..834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1494..2285,2286..2354))
                     /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     2117..2136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2355..2459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2527..2545
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2549..2592)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 aattcttccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 tcctatactt tctagagaat gttttagagc tagaaatagc aagttaaaat aaggctagtc
      121 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgaagctt gggcccgaac
      181 aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat catcatcatt
      241 gagtttaaac ggtctccagc ttggctgttt tggcggatga gagaagattt tcagcctgat
      301 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
      361 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
      421 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
      481 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac tggatcctta
      541 ctcgagtcta gactgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      601 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      661 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      721 gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc
      781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      901 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1141 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2341 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2401 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2461 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2521 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct
     2581 aaggatgatt tctgg
//
