LOCUS       sequence-431018-        5919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5919)
  AUTHORS   Bainton RJ, Kubo KM, Feng JN, Craig NL
  TITLE     Tn7 transposition: target DNA recognition is mediated by multiple 
            Tn7-encoded proteins in a purified in vitro system.
  JOURNAL   Cell 1993 Mar 26;72(6):931-43.
  PUBMED    8384534
REFERENCE   2  (bases 1 to 5919)
  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: "Cell"; 
            date: "1993-03-26"; volume: "72(6)"; pages: "931-43"
COMMENT     Sequence Label: pEMdelta
FEATURES             Location/Qualifiers
     source          1..5919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(108..130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    119..135
                     /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(143..149,150..166,167..172))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(404..1486)
                     /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     1448..1467
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1487..1564)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(1642..1664)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1750..1890
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1805..1824
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1905..1922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2076..2664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2156..2175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2835..3626,3627..3695))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3458..3477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3696..3787)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3806..3833)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3925..4011)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4164..4181
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     mobile_element  4301..4466
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(4587..4606)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4598..5413
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4598..5413
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4671..4690)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tttagagcaa ttcggtgtta gtttcagcaa gcaaacatta accatagcta atgatttata
       61 gccatattaa ccattggggt accgagctcg aattccatgg tctgtttcct gtgtgaaatt
      121 gttatccgct cacaattcca cacattatac gagccggatg attaattgtc aacagctcat
      181 ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc cagaacggga
      241 gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc ataccacagc
      301 ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgatg ataagctgtc
      361 aaaccagatc aattcgcgct aacttacatt aattgcgttg cgctcactgc ccgctttcca
      421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      481 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
      541 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
      601 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
      661 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
      721 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
      781 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
      841 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
      901 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
      961 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     1021 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     1081 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     1141 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     1201 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     1261 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     1321 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     1381 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     1441 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     1501 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg
     1561 tgtcaacgta aatgcatgcc gcttcgcctt cgcgcgcgaa ttgatctgct gcctcgcgcg
     1621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     1681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1741 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     1801 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     1861 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     1921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     2521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2701 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2761 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2821 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2941 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3421 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     3481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3601 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3661 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3721 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3781 aaacaaaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat
     3841 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc
     3901 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     3961 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc
     4021 agttccctac tctcgcatgg ggagacccca cactaccatc ggcgctacgg cgtttcactt
     4081 ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc aaattctgtt
     4141 ttatcagacc gcttctgcgt tctgatttaa tctgtatcag gctgaaaatc ttctctcatc
     4201 cgccaaaaca gccaagcttg catgcctgca ggtcgactct agaggatccc caagaaagtc
     4261 cgtcggacag ctttaataaa ccctgcactt atctgtttag tgtgggcgga caaaatagtt
     4321 gggaactggg aggggtggaa atggagtttt taaggattat ttagggaaga gtgacaaaat
     4381 agatgggaac tgggtgtagc gtcgtaagct aatacgaaaa ttaaaaatga caaaatagtt
     4441 tggaactaga tttcacttat ctggttggtc gacctgcagg gggggggaaa gccacgttgt
     4501 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     4561 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     4621 gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     4681 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     4741 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     4801 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     4861 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
     4921 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4981 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     5041 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     5101 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     5161 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     5221 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     5281 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     5341 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     5401 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     5461 ttgacgggac ggcggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     5521 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     5581 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     5641 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc
     5701 cccccccctg caggtcgacc ccacgcccct ctttaatacg acgggcaatt tgcacttcag
     5761 aaaatgaaga gtttgcttta gccataacaa aagtccagta tgctttttca cagcataact
     5821 ggactgattt cagtttacaa ctattctgtc tagtttaaga ctttattgtc atagtttaga
     5881 tctattttgt tcagtttaag actttattgt ccgcccaca
//
