LOCUS       sequence-473509-        5419 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 5419)
  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 5419)
  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-HP1alpha
FEATURES             Location/Qualifiers
     source          1..5419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(13..18,19..36,37..43)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    51..67
                     /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     56..78
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     75..91
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..91
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        112..130
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    166..300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(889..1602)
                     /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(929..949)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1544..1572
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(1603..1620)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1623..1632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1685..1704)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1717..1735)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1718..1735)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1945..1969)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1966..2169)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1995..2015)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2170..2473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2576..2710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2719..2738)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2720..2738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2745..2762)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2745..2761)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2754..2776)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2903..3358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2990..3009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3200..3221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3384..3488
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3489..3557,3558..4349)
                     /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(3707..3726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4520..5108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5009..5028
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5262..5279
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5396..5417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
       61 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
      121 tcactaaagg gaacaaaagc tggtaccggg ccccccctcg aggtcgatcc agacatgata
      181 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
      241 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
      301 gggttaagct tagctctttg ctgtttcttt ctctttgttt tccgcatcct caggatatgc
      361 atgccatgtc agtctctctt cataaaatgc tatcacaatt tgtggacatt tcacattagc
      421 ttcttttgca agaaccaggt cagcttcatc tgtgtctttc catttcatta ggaacattaa
      481 atcaccacag gaatctgttg ccccaatgat cttttctggt tccagtcctc tctcaaagcc
      541 ccgagcgata tcattgctct gctctctctt ttttttagat ttgatgtcat cggcactgtt
      601 tgagaaattg gatttcctct tgttactttc tgacttctcc ctgggtttat tattttcacc
      661 ctccttcatc ttcttatact ttttcataaa ttcagaaatt agctcagggc aatccaagtt
      721 tttctcaggt tcccaagtat tgtgctcctc agaaaagcct ttccacttca gtagatattc
      781 cacttgtccc ttaaccacgc gcctgtctag caccttctcc acgacatact cctcctcatc
      841 ctctgaagaa gaactgtcag ctgtccgctt ggttttcttt ccggatccct tgtatagttc
      901 atccatgcca tgtgtaatcc cagcagctgt tacaaactca agaaggacca tgtggtctct
      961 cttttcgttg ggatctttcg aaagggcaga ttgtgtggac aggtaatggt tgtctggtaa
     1021 aaggacaggg ccatcgccaa ttggagtatt ttgttgataa tggtctgcta gttgaacgct
     1081 tccatcttca atgttgtgtc taattttgaa gttaactttg attccattct tttgtttgtc
     1141 tgccatgatg tatacattgt gtgagttata gttgtattcc aatttgtgtc caagaatgtt
     1201 tccatcttct ttaaaatcaa taccttttaa ctcgattcta ttaacaaggg tatcaccttc
     1261 aaacttgact tcagcacgtg tcttgtagtt cccgtcatct ttgaaaaata tagttctttc
     1321 ctgtacataa ccttcgggca tggcactctt gaaaaagtca tgccgtttca tatgatctgg
     1381 gtatcttgaa aagcattgaa caccataagt gaaagtagtg acaagtgttg gccatggaac
     1441 aggtagtttt ccagtagtgc aaataaattt aagggtaagt tttccgtatg ttgcatcacc
     1501 ttcaccctct ccactgacag aaaatttgtg cccattaaca tcaccatcta attcaacaag
     1561 aattgggaca actccagtga aaagttcttc tcctttactc atgtgatggt gatggtgatg
     1621 tcccatggtg gcgctagttc tagagcggcc cagaaccgag gtgcagttgg acctgggagt
     1681 ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat
     1741 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt
     1801 gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc
     1861 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
     1921 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     1981 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2041 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2101 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     2161 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     2221 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     2281 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     2341 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     2401 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     2461 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     2521 ctattaataa ctagaggagc atgcgacgtc gggccgccac cgcggtgggg ggatcgatcc
     2581 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2641 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2701 taaacaagtt ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     2761 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2821 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2881 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3301 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     3361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3481 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3601 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3661 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3721 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3781 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3901 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4021 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4081 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4321 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4501 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4561 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4621 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4681 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4741 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4801 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4861 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4921 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4981 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5041 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5101 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5161 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5221 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5281 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5341 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5401 tgagttagct cactcatta
//
