LOCUS       pTmpl-miniDonor-        4490 bp    DNA     circular SYN 07-APR-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4490)
  AUTHORS   grace
  TITLE     Direct Submission
  JOURNAL   Exported Apr 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2283..2288
                     /mol_type="other DNA"
                     /organism="pcDNA3.1(+)"
     promoter        43..147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(148..216,217..1008)
                     /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"
     rep_origin      1179..1767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2055..2076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        join(2091..2096,2097..2114,2115..2121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2129..2145
                     /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)."
     primer_bind     2153..2169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            2183..2282
                     /gene="RNA45SN5"
                     /gene_synonym="RNA45S5"
                     /label=RNA45SN5
                     /note="RNA, 45S pre-ribosomal N5; Derived by automated 
                     computational analysis using gene prediction method: 
                     BestRefSeq."
                     /db_xref="GeneID:100861532"
                     /db_xref="HGNC:HGNC:53530"
     gene            2183..2282
                     /gene="RNA28SN5"
                     /gene_synonym="RN28S1; RNA28S5"
                     /label=RNA28SN5
                     /note="RNA, 28S ribosomal N5; Derived by automated
                     computational analysis using gene prediction method: 
                     BestRefSeq."
                     /db_xref="GeneID:100008589"
                     /db_xref="HGNC:HGNC:53532"
     misc_feature    2183..2282
                     /label=100bp 5' homology for R2 insertion
     misc_feature    2249..2261
                     /label=RUM
                     /note="https://www.science.org/doi/10.1126/science.adg7883
                     "
     misc_feature    2277..2282
                     /label=RAISIN
                     /note="'>https://www.science.org/doi/10.1126/science.adg788
                     3"
     misc_feature    2283..2288
                     /label=BamHI
     promoter        2289..2876
                     /label=CMV promoter
     enhancer        2292..2671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2672..2875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    2815..2833
                     /label=CMV For
     misc_feature    2815..2833
                     /label=CGCCCCATTGACGCAAATG
                     /label=nonstandard type: PCR_primer
     misc_feature    2879..2903
                     /label=TGGCTAACTAGAGAACCCACTGCTT
                     /label=nonstandard type: PCR_primer
     regulatory      2904..2913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2910..3629
                     /codon_start=1
                     /label=GFP ORF
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3630..3678
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     misc_feature    3679..3684
                     /label=HindIII
     misc_feature    3685..3784
                     /label=3' UTR
     gene            3785..3884
                     /gene="RNA45SN5"
                     /gene_synonym="RNA45S5"
                     /label=RNA45SN5
                     /note="RNA, 45S pre-ribosomal N5; Derived by automated 
                     computational analysis using gene prediction method: 
                     BestRefSeq."
                     /db_xref="GeneID:100861532"
                     /db_xref="HGNC:HGNC:53530"
     gene            3785..3884
                     /gene="RNA28SN5"
                     /gene_synonym="RN28S1; RNA28S5"
                     /label=RNA28SN5
                     /note="RNA, 28S ribosomal N5; Derived by automated
                     computational analysis using gene prediction method: 
                     BestRefSeq."
                     /db_xref="GeneID:100008589"
                     /db_xref="HGNC:HGNC:53532"
     misc_feature    3785..3884
                     /label=100bp 3' homology for R2 insertion
     primer_bind     complement(3894..3910)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4052..4480
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 attaacgctt acrmktymsr tkssmcwtty mggsgaaatg tgcgcggaac ccctatttgt
       61 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      121 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      181 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      241 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      301 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      361 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      421 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
      481 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
      541 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
      601 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
      661 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
      721 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
      781 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
      841 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
      901 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
      961 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1021 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1081 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1141 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1201 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1261 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1321 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1381 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1441 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1501 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1561 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1621 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1681 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1741 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1801 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1921 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     1981 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2041 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     2101 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     2161 agctatgacc atgaggcgcg ccgttgacgc gatgtgattt ctgcccagtg ctctgaatgt
     2221 caaagtgaag aaattcaatg aagcgcgggt aaacggcggg agtaactatg actctcttaa
     2281 ggggatccgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2341 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2401 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2461 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2521 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2581 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2641 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2701 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2761 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2821 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2881 gctaactaga gaacccactg cttgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2941 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3001 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3061 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3121 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3181 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3241 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3301 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3361 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3421 acatcgagga cggcagcgtg cagctggccg accactacca gcagaacacc cccatcggcg
     3481 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3541 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3601 ctctcggcat ggacgagctg tacaagtaaa ataaaagatc tttattttca ttagatctgt
     3661 gtgttggttt tttgtgtgaa gcttcacctg atttggacca attcacggga tttgtccaag
     3721 gtggacgggc cacctttact taacccggaa aaggaacata tataatttat gtgtgttcga
     3781 taaatagcca aatgcctcgt catctaatta gtgacgcgca tgaatggatg aacgagattc
     3841 ccactgtccc tacctactat ccagcgaaac cacagccaag ggaagcggcc gccactggcc
     3901 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3961 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4021 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4261 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4381 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
//
