LOCUS       sequence-416615-        5204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GstTnpA from pLac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5204)
  AUTHORS   Meers C, Le HC, Pesari SR, Hoffmann FT, Walker MWG, Gezelle J, Tang 
            S, Sternberg SH
  TITLE     Transposon-encoded nucleases use guide RNAs to promote their selfish
            spread.
  JOURNAL   Nature. 2023 Sep 27. doi: 10.1038/s41586-023-06597-1.
  PUBMED    37758954
REFERENCE   2  (bases 1 to 5204)
  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: "Nature. 
            2023 Sep 27. doi: 10.1038/s41586-023-06597-1."
COMMENT     Sequence Label: pSL4529 (pTnpA)
FEATURES             Location/Qualifiers
     source          1..5204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1193..1384
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1378..1400)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1486..1625
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1541..1560
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1640..1657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1811..2399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1891..1910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2480..2572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2573..2641,2642..3433)
                     /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(2791..2810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3548..4003)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3685..3706)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3897..3916
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4098..4145)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    4616..4632
                     /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)."
     promoter        complement(join(4640..4646,4647..4664,4665..4670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4685..4706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4707..4784
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4785..5204,1..663)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4804..4823)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
       61 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
      121 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
      181 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
      241 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
      301 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
      361 gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga agacagctca
      421 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
      481 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
      541 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
      601 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      661 tgagggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
      721 gcggggcatg actagcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
      781 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
      841 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
      901 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
      961 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     1021 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     1081 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     1141 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     1201 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     1261 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     1321 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     1381 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     1441 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     1501 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     1561 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     1621 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2221 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2461 ggattttggt catgacctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2521 atgagacaat aaccctgata aatgcttcaa tcatgattga aaaaggaaga gtatgagtat
     2581 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2641 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2701 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2761 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2821 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2881 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2941 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3001 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3061 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3121 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3181 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3241 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3301 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3361 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3421 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3481 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgccatcgtg
     3541 ccgccagaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3601 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3661 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3721 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3781 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3841 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3901 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3961 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg
     4021 ttgcgccatt cgatggtgtc cgggcgcatc aggcgccatt cgccattcag gctgcgcaac
     4081 tgttgggaag ggcgatgcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     4141 gctagttatt gctcagcgca ggcagcaagc ctaggttatc atttcatccc ctgattttcg
     4201 atgtattttt tgatgacctc gataggcgct cctcctgttg taagcaggca aaatgagcgg
     4261 gaccaaaagt actccttcca caatttgcgc tttacttgag gaaaatgctt cttaataagg
     4321 cgacttgaag cagacttgta cgcattaatg aacttcgaca attccgagtt ggggtgagct
     4381 ttgaataaga tgtgaacatg gtccatgtca tgattccact ctaccaggct aatgttgtaa
     4441 ttcttaccta aacggacaaa catatccttc gcgtaatccg agattgtgtc atcaatcacc
     4501 tggcgacgat acttgacaac taataccaaa tgatagtaca gaaggaagac gctatggttg
     4561 ttattatcca acttcattcg aagtctcctt cgctcgaagt cggcgcgccg tgaaattgtt
     4621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4681 cctaatgagt gagctaactc acattagaca ccatcgaatg gcgcaaaacc tttcgcggta
     4741 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     4801 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     4861 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     4921 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     4981 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5041 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5101 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5161 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgc
//
