LOCUS       sequence-375154-        7261 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 7261)
  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 7261)
  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: mH2A1.2-CT-GFP
FEATURES             Location/Qualifiers
     source          1..7261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        191..570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        571..774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     725..745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     819..838
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        819..837
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2065..2067,2068..2070,2071..2784)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2098..2126)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2721..2741
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2806..2823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2812..3036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3082..3510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3169..3188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3379..3400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3519..3539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3524..3853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3704..3839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3766..3785
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3920..4714
                     /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(3974..3993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4584..4603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4890..5011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4927..4946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4981..5000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5060..5076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5060..5076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5073..5095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5084..5100
                     /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(5108..5114,5115..5132,5133..5138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5153..5174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5291..5308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5462..6050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5542..5561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6221..7012,7013..7081))
                     /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     6844..6863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7082..7186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7261,1..19))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
       61 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      121 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      181 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      241 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      301 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      361 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      421 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      481 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      541 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      601 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      661 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      721 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      781 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      841 acccaagctg gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
      901 attctcatgt cgagccgcgg tgggaagaag aagtccacca agacgtccag gtctgccaaa
      961 gcaggagtca tctttcccgt ggggcggatg ctgcggtaca tcaagaaagg ccaccccaag
     1021 tacaggattg gagtgggggc acccgtgtac atggccgccg tcctggaata cctgacagcg
     1081 gagattctgg agctggctgg caatgcagcg agagacaaca agaagggacg ggtcacaccc
     1141 cggcacatcc tgctggctgt ggccaatgat gaagagctga atcagctgct aaaaggagtc
     1201 accatagcca gtgggggtgt gttacccaac atccaccccg agttgctagc gaagaagcgg
     1261 ggatccaaag gaaagttgga agccatcatc acaccacccc cagccaaaaa ggccaagtct
     1321 ccatcccaga agaagcctgt atctaaaaaa gcaggaggca agaaaggggc ccggaaatcc
     1381 aagaagcagg gtgaagtcag taaggcagcc agcgccgaca gcacaaccga gggcacacct
     1441 gccgacggct tcacagtcct ctccaccaag agcctcttcc ttggccagaa gctgaacctt
     1501 attcacagtg aaatcagtaa tttagccggc tttgaggtgg aggccataat caatcctacc
     1561 aatgctgaca ttgaccttaa agatgaccta ggaaacacgc tggagaagaa aggtggcaag
     1621 gagtttgtgg aagctgtcct ggaactccgg aaaaagaacg ggcccttgga agtagctgga
     1681 gctgctgtca gcgcaggcca tggcctgcct gccaagtttg tgatccactg taatagtcca
     1741 gtttggggtg cagacaagtg tgaagaactt ctggaaaaga cagtgaaaaa ctgcttggcc
     1801 ctggctgatg ataagaagct gaaatccatt gcatttccat ccatcggcag cggcaggaac
     1861 ggttttccaa agcagacagc agctcagctg attctgaagg ccatctccag ttacttcgtg
     1921 tctacaatgt cctcttccat caaaacggtg tacttcgtgc tttttgacag cgagagtata
     1981 ggcatctatg tgcaggaaat ggccaagctg gacgccaacg aattctgcag atatccagca
     2041 cagtggcggc cgctcgagtc tagaatggct agcaaaggag aagaactttt cactggagtt
     2101 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     2161 gagggtgaag gtgatgctac atacggaaag cttaccctta aatttatttg cactactgga
     2221 aaactacctg ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt
     2281 tcccgttatc cggatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt
     2341 tatgtacagg aacgcactat atctttcaaa gatgacggga actacaagac gcgtgctgaa
     2401 gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaaaggtat tgattttaaa
     2461 gaagatggaa acattctcgg acacaaactc gagtacaact ataactcaca caatgtatac
     2521 atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacatt
     2581 gaagatggat ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     2641 cctgtccttt taccagacaa ccattacctg tcgacacaat ctgccctttc gaaagatccc
     2701 aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat
     2761 ggcatggatg agctctacaa ataatgaatt aaacccgctg atcagcctcg actgtgcctt
     2821 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2881 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2941 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3001 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3061 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3421 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3481 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3601 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3721 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3781 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3841 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3901 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3961 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4021 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4081 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4141 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4201 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4261 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4321 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4381 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4441 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4501 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4561 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4621 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4681 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     4741 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4801 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4861 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4921 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4981 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5041 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5101 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5161 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5221 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5281 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5341 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5401 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5461 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5521 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5581 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5641 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5701 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5761 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5821 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5881 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5941 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6481 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6541 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6601 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6661 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6721 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6781 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6841 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6901 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6961 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7021 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7081 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7141 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7201 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     7261 t
//
