LOCUS       sequence-151862-        6605 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 6605)
  AUTHORS   Chadwick BP, Willard HF
  TITLE     Histone H2A variants and the inactive X chromosome: identification 
            of a second macroH2A variant.
  JOURNAL   Hum Mol Genet. 2001 May 1;10(10):1101-13.
  PUBMED    11331621
REFERENCE   2  (bases 1 to 6605)
  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: "Hum Mol 
            Genet."; date: "2001-05-1"; volume: "10(10)"; pages: "1101-13"
COMMENT     Sequence Label: mH2A1.1-CT-MYC
FEATURES             Location/Qualifiers
     source          1..6605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        126..505
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        506..709
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     660..680
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     754..773
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        754..772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2000..2029
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2045..2062
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2085..2102)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2091..2315
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2361..2789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2448..2467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2658..2679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2798..2818)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2803..3132
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2983..3118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3045..3064
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3199..3993
                     /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(3253..3272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3863..3882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4169..4290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4206..4225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4260..4279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4339..4355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4339..4355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4352..4374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4363..4379
                     /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(4387..4393,4394..4411,4412..4417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4432..4453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4570..4587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4741..5329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4821..4840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5500..6291,6292..6360))
                     /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     6123..6142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6361..6465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6540..6559)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
       61 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      121 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      181 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      361 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      421 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      601 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      661 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      721 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      781 agctggctag ttaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattct
      841 catgtcgagc cgcggtggga agaagaagtc caccaagacg tccaggtctg ccaaagcagg
      901 agtcatcttt cccgtggggc ggatgctgcg gtacatcaag aaaggccacc ccaagtacag
      961 gattggagtg ggggcacccg tgtacatggc cgccgtcctg gaatacctga cagcggagat
     1021 tctggagctg gctggcaatg cagcgagaga caacaagaag ggacgggtca caccccggca
     1081 catcctgctg gctgtggcca atgatgaaga gctgaatcag ctgctaaaag gagtcaccat
     1141 agccagtggg ggtgtgttac ccaacatcca ccccgagttg ctagcgaaga agcggggatc
     1201 caaaggaaag ttggaagcca tcatcacacc acccccagcc aaaaaggcca agtctccatc
     1261 ccagaagaag cctgtatcta aaaaagcagg aggcaagaaa ggggcccgga aatccaagaa
     1321 gcagggtgaa gtcagtaagg cagccagcgc cgacagcaca accgagggca cacctgccga
     1381 cggcttcaca gtcctctcca ccaagagcct cttccttggc cagaagttgc aagttgtaca
     1441 ggctgacatt gcctcgatcg acagtgatgc tgtcgttcac ccgacaaaca ctgacttcta
     1501 catcggtggt gaagtaggaa acacgctgga gaagaaaggt ggcaaggagt ttgtggaagc
     1561 tgtcctggaa ctccggaaaa agaacgggcc cttggaagta gctggagctg ctgtcagcgc
     1621 aggccatggc ctgcctgcca agtttgtgat ccactgtaat agtccagttt ggggtgcaga
     1681 caagtgtgaa gaacttctgg aaaagacagt gaaaaactgc ttggccctgg ctgatgataa
     1741 gaagctgaaa tccattgcat ttccatccat cggcagcggc aggaacggtt ttccaaagca
     1801 gacagcagct cagctgattc tgaaggccat ctccagttac ttcgtgtcta caatgtcctc
     1861 ttccatcaaa acggtgtact tcgtgctttt tgacagcgag agtataggca tctatgtgca
     1921 ggaaatggcc aagctggacg ccaacgaatt ctgcagatat ccagcacagt ggcggccgct
     1981 cgagtctaga gggcccttcg aacaaaaact catctcagaa gaggatctga atatgcatac
     2041 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2101 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2161 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2221 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2281 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2341 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2401 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2461 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2521 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2581 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2641 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2701 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2761 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2881 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2941 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3001 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3061 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3121 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3181 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3241 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3301 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3361 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3421 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3481 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3541 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3601 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3661 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3721 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3781 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3841 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3901 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3961 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     4021 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4081 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4141 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4201 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4261 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4321 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5161 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6481 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     6541 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     6601 gtcgc
//
