LOCUS       sequence-218571-        5527 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 5527)
  AUTHORS   Czlapinski JL, Schelle MW, Miller LW, Laughlin ST, Kohler JJ, 
            Cornish VW, Bertozzi CR
  TITLE     Conditional glycosylation in eukaryotic cells using a biocompatible 
            chemical inducer of dimerization.
  JOURNAL   J Am Chem Soc. 2008 Oct 8. 130(40):13186-7.
  PUBMED    18788807
REFERENCE   2  (bases 1 to 5527)
  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: "J Am Chem 
            Soc. 2008 Oct 8. 130(40):13186-7."
COMMENT     Sequence Label: DHFR-myc
FEATURES             Location/Qualifiers
     source          1..5527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             985..1458
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             1459..1488
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1523..1540)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1529..1753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1799..2227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1886..1905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2096..2117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2236..2256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2241..2570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2421..2556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2483..2502
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2618..2665
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2684..3058
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3188..3309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3225..3244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3279..3298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3358..3374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3358..3374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3371..3393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3382..3398
                     /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(3406..3412,3413..3430,3431..3436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3451..3472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3589..3606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3760..4348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3840..3859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4519..5310,5311..5379))
                     /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     5142..5161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5380..5484)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      841 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa
      901 gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattctgcag atatccagca
      961 cagtggcggc cgctcgagtc tagaatcagt ctgattgcgg cgttagcggt agatcgcgtt
     1021 atcggcatgg aaaacgccat gccgtggaac ctgcctgccg atctcgcctg gtttaaacgc
     1081 aacaccttaa ataaacccgt gattatgggc cgccatacct gggaatcaat cggtcgtccg
     1141 ttgccaggac gcaaaaatat tatcctcagc agtcaaccgg gtacggacga tcgcgtaacg
     1201 tgggtgaagt cggtggatga agccatcgcg gcgtgtggtg acgtaccaga aatcatggtg
     1261 attggcggcg gtcgcgttta tgaacagttc ttgccaaaag cgcaaaaact gtatctgacg
     1321 catatcgacg cagaagtgga aggcgacacc catttcccgg attacgagcc ggatgactgg
     1381 gaatcggtat tcagcgaatt ccacgatgct gatgcgcaga actctcacag ctattgcttt
     1441 gagattctgg agcggcggga acaaaaactc atctcagaag aggatctgta gactagaggg
     1501 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1561 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1621 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1681 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1741 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1861 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1981 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2101 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2221 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2281 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2341 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2401 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2461 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2521 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2581 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     2641 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     2701 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     2761 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     2821 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     2881 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     2941 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     3001 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     3061 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3121 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3181 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3241 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3301 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4201 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     5521 atcggga
//
