LOCUS       sequence-315064-        6106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rhlI and sfCFP (both tagged with ssrA) from the Pcin 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6106)
  TITLE     Bennett Lab Plasmids and Strains
REFERENCE   2  (bases 1 to 6106)
  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"
COMMENT     Sequence Label: pDZ033
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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             219..248
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    433..449
                     /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)."
     terminator      1328..1355
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1604..2419)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSSMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2327..2346
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2411..2430
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2541..3129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3030..3049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3283..3300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3315..3454
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3380..3399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3540..3562
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3556..3747)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(3957..4028)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4225..4938)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     protein_bind    5023..5039
                     /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             complement(5224..5253)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    5455..5471
                     /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)."
ORIGIN
        1 ttgtgagcgg ataacaattt cacacagggc cctcggacac cgaggagaat gtcaagaggc
       61 gaacacacaa cgtcttggag cgccagagga ggaacgagct aaaacggagc ttttttgccc
      121 tgcgtgacca gatcccggag ttggaaaaca atgaaaaggc ccccaaggta gttatcctta
      181 aaaaagccac agcatacatc ctgtccgtcc aagcagagga gcaaaagctc atttctgaag
      241 aggacttgtt gcggaaacga cgagaacagt tgaaacacaa acttgaacag ctacggaact
      301 cttgtgcgta aggaaaagta aggaaaacga ttccttctaa cagaaatgtc ctgagcaatc
      361 acctatgaac tgtcgagggg gcctatctga gggaatttac ttgtgagcgc tcacaattga
      421 tactgagcac aattgtgagc ggataacaat tgcatgccac ttaaaaagga gatcaacaat
      481 gaaagcaatt ttcgtactga aacatcttaa tcatgctaag gagaaatact agtatgatcg
      541 aattgctctc tgaatcgctg gaagggcttt ccgccgccat gatcgccgag ctgggacgct
      601 accggcatca ggtcttcatc gagaagctgg gctgggacgt ggtctccacc tccagggtcc
      661 gcgaccagga attcgaccag ttcgaccatc cgcaaacccg ctacatcgtc gccatgagcc
      721 gccagggcat ctgcggttgc gcccgcctgc tgccgacgac cgacgcctac ctgctcaagg
      781 acgtcttcgc ctacctgtgc agcgaaaccc cgccgagcga tccgtcggtc tgggagcttt
      841 cgcgctacgc cgccagcgcg gcggacgatc cgcagctggc gatgaagata ttctggtcca
      901 gcctgcaatg cgcctggtac ctgggcgcca gttcggtggt ggcggtgacc accacggcca
      961 tggagcgcta tttcgttcgc aacggcgtga tcctccagcg cctcggcccg ccgcagaagg
     1021 tcaagggcga gacgctggtc gcgatcagct tcccggccta ccaggagcgc ggcctggaga
     1081 tgctgctgcg ctaccacccg gaatggctgc agggcgtacc gctgtcgatg gcggtgacct
     1141 ccgcagcgaa cgacgaaaat tacgcccttg cagcgtaata aaagcttccc ctacgtgctg
     1201 ctgaagttgc ccgcaacaga gagtggaacc aaccggtgat accacgatac tatgactgag
     1261 agtcaacgcc atgagcggcc tcatttctta ttctgagtta caacagtccg caccgctgtc
     1321 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagaga gagaatataa
     1381 aaagccagat tattaatccg gcttttttat tatttcctag ctagccgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgacttat
     1501 tcaggcgtag caccaggcgg ctagccgggg aaatgtgcgc ggaaccccta tttgtttatt
     1561 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     1621 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1681 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1741 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1801 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1861 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1921 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     1981 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2041 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2101 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2161 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2221 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2281 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2341 tatacccata taaatcagca tccatgctgg aatttaatcg cggcctagag caagacgttt
     2401 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     2461 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3181 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3241 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3301 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3361 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3421 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3481 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3541 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3601 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3661 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3721 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3781 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3841 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3901 ccagagaaaa atcactcagg gtcaatgcca gcgcttccgc acttatgact ctagtagaga
     3961 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     4021 tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac taccatcggc
     4081 gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg cgctactgcc
     4141 gccaggcaaa ttctgtttta tcagaccgct tctgcgttcg agctcttatt acgctgcaag
     4201 ggcgtaattt tcgtcgttcg ctgctttgta cagttcatcc ataccatgcg tgatgcccgc
     4261 tgcggttacg aactccagca gaaccatatg atcgcgtttc tcgttcggat ctttagacag
     4321 aacgctttgc gtgctcagat agtgattgtc tggcagcaga acaggaccat caccgattgg
     4381 agtgttttgc tggtagtgat cagccagctg cacgctgcca tcctccacgt tgtggcgaat
     4441 tttaaaattc gctttaatgc catttttttg tttatcggcg gtgatgtaaa cattgtcgct
     4501 gatgtaattg tattccagct tatggcccag gatattgccg tcttctttaa agtcaatgcc
     4561 tttcagctca atgcggttta ccagggtatc gccttcaaat ttcacttccg cacgcgtttt
     4621 gtacgtgccg tcatccttaa aggaaatcgt gcgttcctgc acatagcctt ccggcatggc
     4681 ggacttgaag aagtcatgct gcttcatatg gtccggataa cgagcaaagc actgaacacc
     4741 ccaagtcagc gtcgttacca gagtcggcca aggtaccggc agtttaccag tagtacagat
     4801 gaacttcagc gtcagtttac cattagttgc gtcaccttca ccctcgccac gcacggaaaa
     4861 cttatgaccg ttgacatcac catccagttc caccagaata gggacgacac cagtgaacag
     4921 ctcttcgcct ttacgcatac tagtatttct ccttagcatg attaagatgt ttcagtacga
     4981 aaattgcttt cattgttgat ctccttttta agtggcatgc aattgttatc cgctcacaat
     5041 tgtgctcagt atcaattgtg agcgctcaca agtaaattcc ctcagatagg ccccctcgac
     5101 agttcatagg tgattgctca ggacatttct gttagaagga atcgttttcc ttacttttcc
     5161 ttacgcacaa gagttccgta gctgttcaag tttgtgtttc aactgttctc gtcgtttccg
     5221 caacaagtcc tcttcagaaa tgagcttttg ctcctctgct tggacggaca ggatgtatgc
     5281 tgtggctttt ttaaggataa ctaccttggg ggccttttca ttgttttcca actccgggat
     5341 ctggtcacgc agggcaaaaa agctccgttt tagctcgttc ctcctctggc gctccaagac
     5401 gttgtgtgtt cgcctcttga cattctcctc ggtgtccgag ggccctgtgt gaaattgtta
     5461 tccgctcaca attccacaca gacgtcttaa cttgcttcct agtgaaaccg ttcaggacat
     5521 ctacgggatt gttgctaaga aagtcaacga gattctacaa gcagacgcaa tcaatgggac
     5581 cgataacgaa gtagttaccg tgaccgatga gaacactggt gaaatctctg agaaagtcaa
     5641 gctgggcact aaggcactgg ctggtcaatg gctggcttac ggtgttactc gcagtgtgac
     5701 taagcgttca gtcatgacgc tggcttacgg gtccaaagag ttcggcttcc gtcaacaagt
     5761 gctggaagat accattcagc cagctattga ttccggcaag ggtctgatgt tcactcagcc
     5821 gaatcaggct gctggataca tggctaagct gatttgggaa tctgtgagcg tgacggtggt
     5881 agctgcggtt gaagcaatga actggcttaa gtctgctgct aagctgctgg ctgctgaggt
     5941 caaagataag aagactggag agattcttcg caagcgttgc gctgtgcatt gggtaactcc
     6001 tgatggtttc cctgtgtggc aggaatacaa gaagcctatt cagacgcgct tcttcccaac
     6061 cttaccagag ggcgccccag ctggcaattc cgacgtctgt gtggaa
//
