LOCUS       sequence-417782-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 insertion plasmid for Pseudomonas synxantha 2-79. Inserts 
            mNeongreen driven by the J23100 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4274 to 4308)
  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 5729)
  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-J23100-mNeongreen
FEATURES             Location/Qualifiers
     source          1..5729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(93..109)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(94..110)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(524..543)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     643..665
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(703..721)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        789..893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(894..962,963..1754)
                     /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(1112..1131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1925..2513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2414..2433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            2745..2853
                     /note="incP origin of transfer"
     mobile_element  3012..3177
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(3274..3360)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             4239..4250
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(4274..4308)
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     protein_bind    complement(4450..4497)
                     /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(4538..4543,4544..4560,4561..4566)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4755..5288
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(5418..5465)
                     /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(5511..5597)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(join(5700..5729,1..65))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
       61 gagtccaagc tcagctaatt aagctagctt atcgataccg tcgacctcga accccacgcc
      121 cctctttaat acgacgggca atttgcactt cagaaaatga agagtttgct ttagccataa
      181 caaaagtcca gtatgctttt tcacagcata actggactga tttcagttta caactattct
      241 gtctagttta agactttatt gtcatagttt agatctattt tgttcagttt aagactttat
      301 tgtccgccca cacctgctta cgcagggcat ccatttatta ctcaaccgta accgattttg
      361 ccaggttacg cggctggtcc tctagctggc gtaatagcga agaggcccgc accgatcgcc
      421 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
      481 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
      541 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
      601 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
      661 agaggttttc accgtcgtca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
      721 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1861 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2041 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2101 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2161 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2221 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2281 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2341 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2401 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2461 gactcgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2521 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2581 ctgcgttatc aggggattcc ttaaggtata ctttccgctg cataaccctg cttcggggtc
     2641 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     2701 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     2761 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     2821 ctcaacggga atcctgctct gcgaggctgg ccgataagct ctgataccgc tcgccgcagc
     2881 cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2941 ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg cccaaacata acaggaagaa
     3001 aaatgccccg ctgtgggcgg acaaaatagt tgggaactgg gaggggtgga aatggagttt
     3061 ttaaggatta tttagggaag agtgacaaaa tagatgggaa ctgggtgtag cgtcgtaagc
     3121 taatacgaaa attaaaaatg acaaaatagt ttggaactag atttcactta tctggttggc
     3181 ctgcaaggcc ttatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     3241 ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     3301 cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg
     3361 atgcctggca gttccctact ctcgcatggg gagaccccac actaccatcg gcgctacggc
     3421 gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg ccgccaggca
     3481 aattctgttt tatcagaccg cttctgcgtt ctgattcgcg attacttata gagctcatcc
     3541 atccccatca catcggtaaa cgccttctgc cattccttaa aattcagctc cgtcttcgaa
     3601 tgcttgagct cggtcttccg gaacacatac atgggttggt tcttcagata gttcgccgcc
     3661 atcggcttgg caaaggtgta cgtcgtgcgg gccgtcgaac ggtagcgctt gccgttgccg
     3721 gtggtatacg accacttgaa cgtgctgatg atggtcttat cgttcgggta cgtcttcttg
     3781 gaccgacacc aatccgcggc cgtcagggaa ttggtcatca ccggcccgtc cgccggaaag
     3841 ccggtgccct tcacctgcgc ttcccccttg atatggctgc cttcgtacgt gtagcgatag
     3901 ttcacggtca gggaggcgcc atcctcaaac tgcatcgtgc gatgcacttg gtaccccgag
     3961 ccgtcgacca tggccgcctg aaacggggac atcccgtcgg ggtagggcag atattgatgg
     4021 aagccatacc cgatgtgcgg caccaggatc caggggctaa actgcaggtc gcccttcgtg
     4081 ctcttgaggt tgagctcttc gtagccatca ttggggttgc ccgtcccttg ccccaccata
     4141 tcaaagtcca ccccgttgat cgacccgaag atatggagtt catgcgtcgc ggggagggac
     4201 gccatattat cttcctcgcc cttggagacc atgcttaatt tctcctcttt ccgctactag
     4261 taggtctcta gtagctagca ctgtacctag gactgagcta gccgtcaact cctgagaccc
     4321 actagtctac gatgttagcg gcgacaataa tcaccacaca accgaggctc aacaccgtta
     4381 ccggacggcg tcctgcattg ttccactctt tgagctcatg catgatcgaa ttagcttcaa
     4441 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaag
     4501 atcccctgat tccctttgtc aacagcaatg gatcgaattg acataagcct gttcggttcg
     4561 taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     4621 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca
     4681 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     4741 atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta
     4801 aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc
     4861 aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga gacgtagcca
     4921 cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat
     4981 tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc
     5041 tgcccaagtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggcg
     5101 agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg
     5161 cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc
     5221 tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac ccaagtaccg
     5281 ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg ttcggtaaat
     5341 tgtcacaacg ccgcggccaa ttcgatctag aattccattg agtaagtttt taagcacatc
     5401 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaataggt
     5461 acttcaagat ccccaattcg atcgtccggg ccgcaagctc ctagcggcgg atttgtccta
     5521 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga
     5581 gcctttcgtt ttatttgatg cctcaagcta gagagtcatt accccaggcg tttaagggca
     5641 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt ctagcttgga
     5701 ttctcaccaa taaaaaacgc ccggcggca
//
