LOCUS       sequence-424684-        5945 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 5945)
  AUTHORS   Vassallo MF, Tanese N
  TITLE     Isoform-specific interaction of HP1 with human TAFII130.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Apr 30. 99(9):5919-24.
  PUBMED    11959914
REFERENCE   2  (bases 1 to 5945)
  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: "Proc Natl 
            Acad Sci U S A. 2002 Apr 30. 99(9):5919-24."
COMMENT     Sequence Label: pcDNA HA HP1 gamma
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..99)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(253..841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(333..352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1012..1803,1804..1872))
                     /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     1635..1654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1873..1977)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2052..2071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2243..2622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2623..2826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2777..2797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2871..2890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2871..2889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2927..2953
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3506..3524)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3507..3524)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3544..3561)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3550..3774
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3820..4248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4117..4138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4257..4277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4262..4591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4442..4577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4504..4523
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4658..5452
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4712..4731)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5322..5341
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5626..5747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5663..5682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5717..5736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5796..5812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5796..5812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5809..5831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5820..5836
                     /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(5844..5850,5851..5868,5869..5874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5889..5910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
       61 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
      721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1981 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     2041 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     2101 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     2161 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     2221 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     2281 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2341 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2401 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2461 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2521 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2581 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2641 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2701 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2761 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2821 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     2881 cactataggg agacccaagc ttggtaccga gctcggatcc accatgtacc catacgatgt
     2941 tccagattac gctgaattcg gaaaaaaaca gaatggaaag agtaaaaaag ttgaagaggc
     3001 agagcctgaa gaatttgtcg tggaaaaagt actagatcga cgtgtagtga atgggaaagt
     3061 ggaatatttc ctgaagtgga agggatttac agatgctgac aatacttggg aacctgaaga
     3121 aaatttagat tgtccagaat tgattgaagc gtttcttaac tctcagaaag ctggcaaaga
     3181 aaaagatggt acaaaaagaa aatctttatc tgacagtgaa tctgatgaca gcaaatcaaa
     3241 gaagaaaaga gatgctgctg acaaaccaag aggatttgcc agaggtcttg atcctgaaag
     3301 aataattggt gccacagaca gcagtggaga attgatgttt ctcatgaaat ggaaagattc
     3361 agatgaggca gacttggtgc tggcgaaaga ggcaaatatg aagtgtcctc aaattgtaat
     3421 tgctttttat gaagagagac taacttggca ttcttgtcca gaagatgaag ctcaataact
     3481 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     3541 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3601 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3661 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3721 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3781 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4021 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4141 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4201 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4261 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4321 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4441 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4501 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4561 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4621 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4681 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4741 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4801 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4861 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4921 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4981 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5041 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5101 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5161 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5221 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5281 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5341 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5401 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5461 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5521 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5581 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5761 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5821 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5881 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5941 tcggg
//
