LOCUS       sequence-397577-        2810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of eukaryotic genes of interest with CMV promoter 
            with EcoRI as a PlayBack Restriction Enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2810)
  AUTHORS   Foran G, Hallam RD, Megaly M, Turgambayeva A, Necakov A
  TITLE     PlayBack cloning: simple, reversible, cost-effective cloning for the
            combinatorial assembly of complex expression constructs.
  JOURNAL   Biotechniques. 2023 Oct;75(4):168-178. doi: 10.2144/btn-2023-0042. 
            Epub 2023 Oct 10.
  PUBMED    37815818
REFERENCE   2  (bases 1 to 2810)
  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: 
            "10.2144/btn-2023-0042"; journalName: "Biotechniques"; date: 
            "2023-10"; volume: "75"; issue: "4"; pages: "168-178"
COMMENT     Sequence Label: PlayBack-CMV-EcoR1
FEATURES             Location/Qualifiers
     source          1..2810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    778..1002
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(1073..1276)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1102..1122)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1277..1580
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1675..1779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1780..1848,1849..2640)
                     /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(1998..2017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtat ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagccat ggtcgaccag aattcgacca
      781 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
      841 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
      901 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      961 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcattcat tcacaagctt
     1021 agtcaatcaa tctagactga gatccctagc ggatctgacg gttcactaaa ccagctctgc
     1081 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     1141 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     1201 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     1261 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     1321 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     1381 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     1441 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     1501 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     1561 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     1621 ttgattacta ttaataacga attcctcgag ttacgcgtaa ttcggggaaa tgtgcgcgga
     1681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
