LOCUS       sequence-232353-        6170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial tdTomato labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6170)
  AUTHORS   Kang YS, Kirby JE
  TITLE     Promotion and Rescue of Intracellular Brucella neotomae Replication 
            during Coinfection with Legionella pneumophila.
  JOURNAL   Infect Immun. 2017 Apr 21;85(5). pii: e00991-16. doi: 
            10.1128/IAI.00991-16. Print 2017 May.
  PUBMED    28264909
REFERENCE   2  (bases 1 to 6170)
  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: "Infect 
            Immun."; date: "2017-04-21"; volume: "85(5). pii: e00991-16. doi"; 
            pages: " 10.1128/IAI.00991-16. Print 2017 May"
COMMENT     Sequence Label: pMOD3-proD/tdTomato-hph
FEATURES             Location/Qualifiers
     source          1..6170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    309..327
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     mobile_element  504..1271
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             2764..3789
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2764..3789
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    4096..4114
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(4180..4197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4351..4939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4431..4450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5110..5901,5902..5970))
                     /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     5733..5752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5971..6075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6143..6161
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgacagct gtctcttata cacatctcaa ccatcatcga tgaattttct cgggtgttct
      361 cgcatattgg ctcgaattcc acagctaaca ccacgtcgtc cctatctgtt gccctaggtc
      421 tatgagtggt tgctggataa ctttacgggc atgcataagg ctcgtataat atattcaggg
      481 agaccacaac ggtttccctc tacggtgatg ctgccaactt actgatttag tgtatgatgg
      541 tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact
      601 gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctatct ctgctctcac
      661 tgccgtaaaa catggcaact gcagttcact tacaccgctt ctcaacccgg tacgcaccag
      721 aaaatcattg atatggccat gaatggcgtt ggatgccggg caacagcccg cattatgggc
      781 gttggcctca acacgatttt acgtcactta aaaaactcag gccgcagtcg gtaacctcgc
      841 gcatacagcc gggcagtgac gtcatcgtct gcgcggaaat ggacgaacag tggggctatg
      901 tcggggctaa atcgcgccag cgctggctgt tttacgcgta tgacagtctc cggaagacgg
      961 ttgttgcgca cgtattcggt gaacgcacta tggcgacgct ggggcgtctt atgagcctgc
     1021 tgtcaccctt tgacgtggtg atatggatga cggatggctg gccgctgtat gaatcccgcc
     1081 tgaagggaaa gctgcacgta atcagcaagc gatatacgca gcgaattgag cggcataacc
     1141 tgaatctgag gcagcacctg gcacggctgg gacggaagtc gctgtcgttc tcaaaatcgg
     1201 tggagctgca tgacaaagtc atcgggcatt atctgaacat aaaacactat caataagttg
     1261 gagtcattac ctccctctac aaataatttt gtttaacttt taaagaggaa aaaaaatggt
     1321 gagtaagggc gaggaggtca ttaaggagtt catgcggttc aaggtcagga tggagggcag
     1381 catgaacggt catgagttcg aaatcgaagg agaaggagag ggccggccgt acgaggggac
     1441 gcagaccgcg aagctgaagg tgacgaaggg cggcccactc cccttcgcat gggacatcct
     1501 gtcgccgcag ttcatgtatg gttccaaggc gtacgtgaag catccggcgg acataccgga
     1561 ctacaagaag ctgagcttcc cggaggggtt caagtgggag cgggtcatga actttgagga
     1621 cggcggcctt gtcacagtga cccaggatag cagtttgcag gacggcacgc tgatctataa
     1681 ggtgaagatg cgagggacca acttcccgcc cgacggcccc gtgatgcaga agaagaccat
     1741 gggctgggag gcttcgacgg agcgcctgta cccgcgtgac ggtgtgctga agggcgaaat
     1801 ccaccaggcc ctgaagctga aggatggagg tcactacctc gtcgagttca agaccatcta
     1861 catggccaaa aagcccgtgc agctccccgg ctactactat gttgacacca agctcgacat
     1921 tacctcgcat aacgaggact ataccatcgt ggagcaatac gaacgctcgg agggccggca
     1981 ccacctgttc ctcggtcatg ggaccggttc gactggcagc ggtagttcag gaaccgcctc
     2041 gtcggaggac aacaatatgg ccgtgatcaa agagttcatg cgcttcaagg tgcgaatgga
     2101 aggttcgatg aacggccacg agttcgagat cgagggggaa ggcgaaggtc ggccatacga
     2161 aggcacccag acggcgaagt tgaaagtgac caagggcggc ccgctgccgt tcgcgtggga
     2221 catcctgtca ccgcaattca tgtatggctc gaaggcctac gtcaagcacc cggccgacat
     2281 tccagactac aagaagctct cttttcccga gggcttcaag tgggagcgcg tcatgaactt
     2341 tgaggatggt ggcctggtga ccgtaacgca ggacagctcc ctccaggacg ggaccctgat
     2401 ctataaagtg aagatgcgcg gcaccaactt cccacccgat ggcccagtaa tgcaaaagaa
     2461 aacgatggga tgggaggcaa gcaccgaacg tctgtacccg cgcgacggag tcctgaaggg
     2521 tgaaatccac caagccctaa aactgaagga cggtgggcac tacctggttg agttcaagac
     2581 catctatatg gcaaagaagc ccgtgcaact ccccggatat tactacgtgg acacgaagct
     2641 ggacatcacc agccacaacg aagactacac gatcgtcgag caatacgaac ggagtgaggg
     2701 tcggcaccat ctcttcctct atggtatgga cgagctgtac aagtaagtcg acggaggaca
     2761 gtaatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     2821 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     2881 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     2941 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     3001 attggggaat ttagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     3061 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc aaccggtcgc ggaggccatg
     3121 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     3181 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     3241 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     3301 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     3361 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     3421 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     3481 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     3541 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     3601 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     3661 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     3721 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     3781 aaggaataaa agctttaaaa gccttatata ttcttttttt tcttataaaa cttaaaacct
     3841 tagaggctat ttaagttgct gatttatatt aattttattg ttcaaacatg agagcttagt
     3901 acgtgaaaca tgagagctta gtacgttagc catgagagct tagtacgtta gccatgaggg
     3961 tttagttcgt taaacatgag agcttagtac gttaaacatg agagcttagt acgtgaaaca
     4021 tgagagctta gtacgtacta tcaacaggtt gaactgccaa cgactacgca ctagccaaca
     4081 agagcttcag ggttgagatg tgtataagag acagctgtct taatgaatcg gccaacgcgc
     4141 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4261 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4321 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4861 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4921 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4981 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5041 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5221 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5401 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5941 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6001 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6061 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6121 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
