LOCUS       sequence-181020-        5401 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 5401)
  AUTHORS   Cheutin T, McNairn AJ, Jenuwein T, Gilbert DM, Singh PB, Misteli T
  TITLE     Maintenance of stable heterochromatin domains by dynamic HP1 
            binding.
  JOURNAL   Science. 2003 Jan 31. 299(5607):721-5.
  PUBMED    12560555
REFERENCE   2  (bases 1 to 5401)
  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: "Science. 
            2003 Jan 31. 299(5607):721-5."
COMMENT     Sequence Label: GFP-HP1beta
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        90..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      931..940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             943..960
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             961..1674
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(991..1019)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1614..1634
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2245..2379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2415..2433)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2454..2470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2454..2470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2467..2489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2478..2494
                     /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(2502..2508,2509..2526,2527..2532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2547..2568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2685..2702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2856..3444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2936..2955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3615..4406,4407..4475))
                     /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     4238..4257
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4476..4580)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4606..5061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4743..4764)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4955..4974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5188..5210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5202..5219
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5203..5219
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5226..5245
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5226..5244
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5254..5388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cggtggcggc ccgacgtcgc atgctcctct agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tgggccgctc tagaactagc gccaccatgg gacatcacca tcaccatcac
      961 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     1021 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
     1081 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     1141 gtcactactt tcacttatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg
     1201 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     1261 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1321 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
     1381 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     1441 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     1501 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     1561 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     1621 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caagggatcc
     1681 gggaaaaaac aaaacaagaa gaaagtggag gaggtgctag aagaggagga agaggaatat
     1741 gtggtggaaa aagttctcga ccgtcgagtg gtaaagggca aagtggagta cctcctaaag
     1801 tggaagggat tctcagatga ggacaacaca tgggagccag aagagaacct ggattgcccc
     1861 gacctcattg ctgagtttct gcagtcacag aaaacagcac atgagacaga taaatcagag
     1921 ggaggcaagc gcaaagctga ttctgattct gaagataagg gagaggagag caaaccaaag
     1981 aagaagaaag aagagtcaga aaagccacga ggctttgctc gaggtttgga gccggagcgg
     2041 attattggag ctacagactc cagtggagag ctcatgttcc tgatgaaatg gaaaaactct
     2101 gatgaggctg acctggtccc tgccaaggaa gccaatgtca agtgcccaca ggttgtcata
     2161 tccttctatg aggaaaggct gacgtggcat tcctacccct cggaggatga tgacaaaaaa
     2221 gatgacaaga actaagctta acccaacttg tttattgcag cttataatgg ttacaaataa
     2281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2341 ttgtccaaac tcatcaatgt atcttatcat gtctggatcg acctcgaggg ggggcccggt
     2401 accagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     2461 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2521 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2581 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2641 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2701 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2761 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2821 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2881 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2941 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3001 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3061 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3121 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3181 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3241 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3301 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3361 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3421 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3481 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3541 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3601 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3661 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3721 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3781 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3841 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3901 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3961 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4021 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4081 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4141 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4201 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4261 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4321 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4381 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4441 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4501 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4561 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     4621 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4681 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4741 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4801 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4861 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4921 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4981 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5041 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     5101 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5161 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5221 aattgtaata cgactcacta tagggcgaat tggaacttgt ttattgcagc ttataatggt
     5281 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5341 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcga tccccccacc
     5401 g
//
