LOCUS       sequence-417780-        5706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 insertion plasmid for Pseudomonas synxantha 2-79. Inserts 
            mNeongreen driven by the Syn35 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5706)
  AUTHORS   Meyerowitz JT, Larsson EM, Murray RM
  TITLE     Development of Cell-Free Transcription-Translation Systems in Three 
            Soil Pseudomonads.
  JOURNAL   ACS Synth Biol. 2024 Feb 16;13(2):530-537. doi: 
            10.1021/acssynbio.3c00468. Epub 2024 Feb 6.
  PUBMED    38319019
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: 
            "10.1021/acssynbio.3c00468"; journalName: "ACS Synth Biol"; date: 
            "2024-02-16- 16"; volume: "13"; issue: "2"; pages: "530-537"
COMMENT     Sequence Label: pJM220-Syn35-mNeongreen
FEATURES             Location/Qualifiers
     source          1..5706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      231..819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     720..739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            1051..1159
                     /note="incP origin of transfer"
     mobile_element  1318..1483
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(1580..1666)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2545..2556
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(2733..2780)
                     /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)."
     promoter        join(2821..2826,2827..2843,2844..2849)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3038..3571
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(3701..3748)
                     /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(3794..3880)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3983..4077)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4105..4121)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4106..4122)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4536..4555)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4655..4677
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4715..4733)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4801..4905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4906..4974,4975..5706,1..60)
                     /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(5124..5143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
       61 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
      121 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
      181 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      241 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      301 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      361 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      421 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      481 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
      541 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
      601 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
      661 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      721 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact cgagcgtcga
      781 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
      841 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcaggg
      901 gattccttaa ggtatacttt ccgctgcata accctgcttc ggggtcatta tagcgatttt
      961 ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga
     1021 ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc
     1081 gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc
     1141 tgctctgcga ggctggccga taagctctga taccgctcgc cgcagccgaa cgaccgagcg
     1201 cagcgggtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1261 gcgttggccg attcattaat gcagcgccca aacataacag gaagaaaaat gccccgctgt
     1321 gggcggacaa aatagttggg aactgggagg ggtggaaatg gagtttttaa ggattattta
     1381 gggaagagtg acaaaataga tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta
     1441 aaaatgacaa aatagtttgg aactagattt cacttatctg gttggcctgc aaggccttat
     1501 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
     1561 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     1621 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
     1681 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
     1741 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
     1801 agaccgcttc tgcgttctga ttcgcgatta cttatagagc tcatccatcc ccatcacatc
     1861 ggtaaacgcc ttctgccatt ccttaaaatt cagctccgtc ttcgaatgct tgagctcggt
     1921 cttccggaac acatacatgg gttggttctt cagatagttc gccgccatcg gcttggcaaa
     1981 ggtgtacgtc gtgcgggccg tcgaacggta gcgcttgccg ttgccggtgg tatacgacca
     2041 cttgaacgtg ctgatgatgg tcttatcgtt cgggtacgtc ttcttggacc gacaccaatc
     2101 cgcggccgtc agggaattgg tcatcaccgg cccgtccgcc ggaaagccgg tgcccttcac
     2161 ctgcgcttcc cccttgatat ggctgccttc gtacgtgtag cgatagttca cggtcaggga
     2221 ggcgccatcc tcaaactgca tcgtgcgatg cacttggtac cccgagccgt cgaccatggc
     2281 cgcctgaaac ggggacatcc cgtcggggta gggcagatat tgatggaagc catacccgat
     2341 gtgcggcacc aggatccagg ggctaaactg caggtcgccc ttcgtgctct tgaggttgag
     2401 ctcttcgtag ccatcattgg ggttgcccgt cccttgcccc accatatcaa agtccacccc
     2461 gttgatcgac ccgaagatat ggagttcatg cgtcgcgggg agggacgcca tattatcttc
     2521 ctcgcccttg gagaccatgc ttaatttctc ctctttccta ggtccccaaa ttataattct
     2581 aaacatcacg catgtcaaat aaattaatta aacgatgtta gcggcgacaa taatcaccac
     2641 acaaccgagg ctcaacaccg ttaccggacg gcgtcctgca ttgttccact ctttgagctc
     2701 atgcatgatc gaattagctt caaaagcgct ctgaagttcc tatactttct agagaatagg
     2761 aacttcggaa taggaacttc aagatcccct gattcccttt gtcaacagca atggatcgaa
     2821 ttgacataag cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac
     2881 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     2941 tgttatgact gtttttttgt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     3001 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt
     3061 tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg
     3121 cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg
     3181 tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg attacctcgg
     3241 gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt
     3301 tggcgctctc gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg agatctatat
     3361 ctatgatctc gcagtctccg gcgagcaccg gaggcagggc attgccaccg cgctcatcaa
     3421 tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta
     3481 cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca
     3541 ctttgatatc gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc
     3601 ggccgcggag ttgttcggta aattgtcaca acgccgcggc caattcgatc tagaattcca
     3661 ttgagtaagt ttttaagcac atcagcttca aaagcgctct gaagttccta tactttctag
     3721 agaataggaa cttcggaata ggtacttcaa gatccccaat tcgatcgtcc gggccgcaag
     3781 ctcctagcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     3841 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctcaag ctagagagtc
     3901 attaccccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     3961 cactcatcgc agtctagctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     4021 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     4081 gctcagctaa ttaagctagc ttatcgatac cgtcgacctc gaaccccacg cccctcttta
     4141 atacgacggg caatttgcac ttcagaaaat gaagagtttg ctttagccat aacaaaagtc
     4201 cagtatgctt tttcacagca taactggact gatttcagtt tacaactatt ctgtctagtt
     4261 taagacttta ttgtcatagt ttagatctat tttgttcagt ttaagacttt attgtccgcc
     4321 cacacctgct tacgcagggc atccatttat tactcaaccg taaccgattt tgccaggtta
     4381 cgcggctggt cctctagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4441 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     4501 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4561 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4621 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4681 tcaccgtcgt caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4741 gttaatgtca tgataataac ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4801 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4861 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4921 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4981 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5041 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5101 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5161 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5221 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5281 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5341 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5401 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5461 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5521 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5581 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5641 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5701 gcaact
//
