LOCUS       sequence-302079-        7252 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7252)
  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 7252)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-GFP
FEATURES             Location/Qualifiers
     source          1..7252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        516..719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     670..690
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     764..783
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        764..782
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2001..2003,2004..2006,2007..2720)
                     /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(2034..2062)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2657..2677
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2742..2759)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2748..2972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3018..3446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3105..3124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3315..3336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3455..3475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3460..3789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3640..3775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3702..3721
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3856..4650
                     /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(3910..3929)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4520..4539
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4826..4947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4863..4882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4917..4936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4996..5012)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4996..5012)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5009..5031)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5020..5036
                     /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(5044..5050,5051..5068,5069..5074))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5089..5110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5227..5244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5398..5986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5478..5497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6157..6948,6949..7017))
                     /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     6780..6799
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7018..7122)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7197..7216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
       61 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      121 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      481 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      541 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      601 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      661 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      721 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
      781 gggagaccca agctggctag ttaagcttgg taccgagctc ggatccacta gtccagtgtg
      841 gtggaattct catgtcgagc cgcggtggga agaagaagtc caccaagacg tccaggtctg
      901 ccaaagcagg agtcatcttt cccgtggggc ggatgctgcg gtacatcaag aaaggccacc
      961 ccaagtacag gattggagtg ggggcacccg tgtacatggc cgccgtcctg gaatacctga
     1021 cagcggagat tctggagctg gctggcaatg cagcgagaga caacaagaag ggacgggtca
     1081 caccccggca catcctgctg gctgtggcca atgatgaaga gctgaatcag ctgctaaaag
     1141 gagtcaccat agccagtggg ggtgtgttac ccaacatcca ccccgagttg ctagcgaaga
     1201 agcggggatc caaaggaaag ttggaagcca tcatcacacc acccccagcc aaaaaggcca
     1261 agtctccatc ccagaagaag cctgtatcta aaaaagcagg aggcaagaaa ggggcccgga
     1321 aatccaagaa gcagggtgaa gtcagtaagg cagccagcgc cgacagcaca accgagggca
     1381 cacctgccga cggcttcaca gtcctctcca ccaagagcct cttccttggc cagaagttgc
     1441 aagttgtaca ggctgacatt gcctcgatcg acagtgatgc tgtcgttcac ccgacaaaca
     1501 ctgacttcta catcggtggt gaagtaggaa acacgctgga gaagaaaggt ggcaaggagt
     1561 ttgtggaagc tgtcctggaa ctccggaaaa agaacgggcc cttggaagta gctggagctg
     1621 ctgtcagcgc aggccatggc ctgcctgcca agtttgtgat ccactgtaat agtccagttt
     1681 ggggtgcaga caagtgtgaa gaacttctgg aaaagacagt gaaaaactgc ttggccctgg
     1741 ctgatgataa gaagctgaaa tccattgcat ttccatccat cggcagcggc aggaacggtt
     1801 ttccaaagca gacagcagct cagctgattc tgaaggccat ctccagttac ttcgtgtcta
     1861 caatgtcctc ttccatcaaa acggtgtact tcgtgctttt tgacagcgag agtataggca
     1921 tctatgtgca ggaaatggcc aagctggacg ccaacgaatt ctgcagatat ccagcacagt
     1981 ggcggccgct cgagtctaga atggctagca aaggagaaga acttttcact ggagttgtcc
     2041 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     2101 gtgaaggtga tgctacatac ggaaagctta cccttaaatt tatttgcact actggaaaac
     2161 tacctgttcc atggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttccc
     2221 gttatccgga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     2281 tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca
     2341 agtttgaagg tgataccctt gttaatcgta tcgagttaaa aggtattgat tttaaagaag
     2401 atggaaacat tctcggacac aaactcgagt acaactataa ctcacacaat gtatacatca
     2461 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag
     2521 atggatccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     2581 tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg
     2641 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
     2701 tggatgagct ctacaaataa tgaattaaac ccgctgatca gcctcgactg tgccttctag
     2761 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2821 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2881 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2941 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3001 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3061 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3121 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3181 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3241 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3301 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3361 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3421 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3661 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3721 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3781 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3841 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3901 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3961 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4021 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4081 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4141 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4201 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4261 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4321 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4381 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4441 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4501 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4561 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4621 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4801 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4861 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4921 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4981 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5041 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5101 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5161 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5221 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5281 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5341 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5401 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5461 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5521 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5581 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5641 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5701 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5761 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5821 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5881 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5941 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6001 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6061 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6121 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6181 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6241 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6301 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6361 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6421 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6481 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6541 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6601 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6661 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6721 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6781 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6841 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6901 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6961 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7021 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7081 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7141 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     7201 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gt
//
