LOCUS       sequence-338926-        6420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA donor for transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  AUTHORS   Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, 
            Yang S
  TITLE     Programming Cells by Multicopy Chromosomal Integration Using 
            CRISPR-Associated Transposases.
  JOURNAL   CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018.
  PUBMED    34152213
REFERENCE   2  (bases 1 to 6420)
  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: 
            "10.1089/crispr.2021"; journalName: "CRISPR J"; date: "2021-06"; 
            volume: "4"; issue: "3"; pages: "350-359"
COMMENT     Sequence Label: pRE57I-GNAc
FEATURES             Location/Qualifiers
     source          1..6420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        233..251
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    252..276
                     /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)."
     RBS             291..313
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        2170..2188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2189..2213
                     /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)."
     RBS             2228..2250
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2816..2864
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      2926..2974
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(3043..3065)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      complement(3504..4092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3584..3603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4263..5054,5055..5123))
                     /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     4886..4905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5124..5228)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5259..6341)
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6303..6322
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6342..6419)
                     /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."
ORIGIN
        1 ccaatgatgc gaaaaaaccc cgccgaagcg gggttttttg cgaaaaaaat agggataaca
       61 gggtaatgtt taaactgttg atacaaccat aaaatgataa ttacacccat aaattgataa
      121 ttatcacacc cacgggaatt ccatggtcga cccagcaacc gcacctgtgg cgccggtgat
      181 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
      241 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      301 tttaagaagg agatatacca tgtgtggaat tgttggcgcg atcgcgcaac gtgatgtagc
      361 agaaatcctt cttgaaggtt tacgtcgtct ggaataccgc ggatatgact ctgccggtct
      421 ggccgttgtt gatacagaag gtcatatgac ccgcctgcgt cgcctcggta aagtccagat
      481 gctggcacag gcagcggaag aacatcctct gcatggcggc actggtattg ctcacactcg
      541 ctgggcgacc cacggtgaac cttcagaagt gaatgcgcat ccgcatgttt ctgaacacat
      601 tgtggtggtg cataacggca tcatcgaaaa ccatgaaccg ctgcgtgaag agctaaaagc
      661 gcgtggctat accttcgttt ctgaaaccga caccgaagtg attgcccatc tggtgaactg
      721 ggagctgaaa caaggcggga ctctgcgtga ggccgttctg cgtgctatcc cgcagctgcg
      781 tggtgcgtac ggtacagtga tcatggactc ccgtcacccg gataccctgc tggcggcacg
      841 ttctggtagt ccgctggtga ttggcctggg gatgggcgaa aactttatcg cttctgacca
      901 gctggcgctg ttgccggtga cccgtcgctt tatcttcctt gaagagggcg atattgcgga
      961 aatcactcgc cgttcggtaa acatcttcga taaaactggc gcggaagtaa aacgtcagga
     1021 tatcgaatcc aatctgcaat atgacgcggg cgataaaggc atttactgtc actacatgca
     1081 gaaagagatc tacgaacagc cgaacgcgat caaaaacacc cttaccggac gcatcagcca
     1141 cggtcaggtt gatttaagcg agctgggacc gaacgccgac gaactgctgt cgaaggttga
     1201 gcatattcag atcctcgcct gtggtacttc ttataactcc ggtatggttt cccgctactg
     1261 gtttgaatcg ctagcaggta ttccgtgcga cgtcgaaatc gcctctgaat tccgctatcg
     1321 caaatctgcc gtgcgtcgta acagcctgat gatcaccttg tcacagtctg gcgaaaccgc
     1381 ggataccctg gctggcctgc gtctgtcgaa agagctgggt taccttggtt cactggcaat
     1441 ctgtaacgtt ccgggttctt ctctggtgcg cgaatccgat ctggcgctaa tgaccaacgc
     1501 gggtacagaa atcggcgtgg catccactaa agcattcacc actcagttaa ctgtgctgtt
     1561 gatgctggtg gcgaagctgt ctcgcctgaa aggtctggat gcctccattg aacatgacat
     1621 cgtgcatggt ctgcaggcgc tgccgagccg tattgagcag atgctgtctc aggacaaacg
     1681 cattgaagcg ctggcagaag atttctctga caaacatcac gcgctgttcc tgagccgtgg
     1741 cgatcagtac ccaatcgcgc tggaaggcgc attgaagttg aaagagatct cttacattca
     1801 cgctgaagcc tacgctgctg gcgaactgaa acacggtccg ctggcgctaa ttgatgccga
     1861 tatgccggtt attgttgttg caccgaacaa cgaattgctg gaaaaactga aatccaacat
     1921 tgaagaagtt cgcgcgcgtg gcggtcagtt gtatgtcttc gccgatcagg atgcgggttt
     1981 tgtaagtagc gataacatgc acatcatcga gatgccgcat gtggaagagg tgattgcacc
     2041 gatcttctac accgttccgc tgcagctgct ggcttaccat gtcgcgctga tcaaaggcac
     2101 cgacgttgac cagccgcgta acctggcaaa atcggttacg gttgagtaag gatctcgatc
     2161 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     2221 aaataatttt gtttaacttt aagaaggaga tataccatga gcttacccga tggattttat
     2281 ataaggcgaa tggaagaggg agatttggaa caggtcactg agacgctaaa ggttttgacc
     2341 accgtgggca ctattacccc cgaatccttc agcaaactca taaaatactg gaatgaagcc
     2401 acagtatgga atgataacga agataaaaaa ataatgcaat ataaccccat ggtgattgtg
     2461 gacaagcgca ccgagacggt tgccgctacg gggaatatca tcatcgaaag aaagatcatt
     2521 catgaactgg ggctatgtgg ccacatcgag gacattgcag taaactccaa gtatcagggc
     2581 caaggtttgg gcaagctctt gattgatcaa ttggtaacta tcggctttga ctacggttgt
     2641 tataagatta ttttagattg cgatgagaaa aatgtcaaat tctatgaaaa atgtgggttt
     2701 agcaacgcag gcgtggaaat gcaaattaga aaatagctcg agttaattaa actagtgagc
     2761 tcggtacccg gggatcctct agaggtcgaa attcacctcg aaagcaagct gataaaccga
     2821 tacaattaaa ggctcctttt ggagcctttt tttttggaga ttttcaacgt gaaaaaatta
     2881 ttattcgcaa ttccaagcta attcacctcg aaagcaagct gataaaccga tacaattaaa
     2941 ggctcctttt ggagcctttt tttttggaga ttttcaacgt gaaaaaatta ttattcgcaa
     3001 ttccaagctc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3061 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3121 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     3181 cggagtgtat gggctcgatc ccctcggatc caattggtcc ttactgcagt agttttgctg
     3241 aaatactcga ttcacaaaaa tatcaactta tggttgtttt gtgagatatc aatatatggt
     3301 tgttttgtgg ttaagttgct gattataaat aattattaaa tatcacttta tggttgcatc
     3361 aacagttaac attaccctgt tatccctaaa aaaaaaaccc cgcccctgac agggcggggt
     3421 ttttttttta atatcggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3901 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4261 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4321 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4381 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4441 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4501 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4561 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4621 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4681 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4741 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4801 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4861 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4921 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4981 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5041 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5221 gttccgcgca catttccccg aaaagtgcca cctgacgttc actgcccgct ttccagtcgg
     5281 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5341 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     5401 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     5461 cgaaaatcct gtttgatggt ggttgacggc gggatataac atgagctgtc ttcggtatcg
     5521 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5581 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5641 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5701 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5761 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5821 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5881 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5941 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     6001 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     6061 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     6121 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     6181 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     6241 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     6301 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     6361 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcc
//
