LOCUS       sequence-461577-        5101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes a H2B-iRFP720 fusion contruct under the control of a dox 
            inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5101)
  AUTHORS   Pelosse M, Marcia M
  TITLE     biGMamAct: efficient CRISPR/Cas9-mediated docking of large 
            functional DNA cargoes at the ACTB locus
  JOURNAL   Synthetic Biology 2025.02.13
REFERENCE   2  (bases 1 to 5101)
  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: "Synthetic 
            Biology 2025.02.13"
COMMENT     Sequence Label: psLIB TRE H2B iRFP720
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      123..711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     612..631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     865..882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1016..1240
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1307..1840)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2029..2034,2035..2051,2052..2057))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2453..2486)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2566..2584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2603..2621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2640..2658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2677..2695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2751..2769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2788..2806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2825..2843
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2865..2889
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3031..3408
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      3421..3430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4570..4704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4607..4626)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4661..4680
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4814..4979
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa tggcgaatgg
       61 gacgcgcccg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      241 ttcagcagag cgcagatacc aaatactatt cttctagtgt agccgtagtt aggccaccac
      301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      781 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      841 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
      901 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt
      961 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg
     1021 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     1081 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     1141 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     1201 aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac
     1261 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     1321 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     1381 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     1441 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     1501 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     1561 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     1621 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     1681 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     1741 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     1801 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     1861 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     1921 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     1981 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     2041 gaacaggctt atgtcaactg ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     2101 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     2161 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     2221 ctgtgcacgg atctgccctt gcttcaggag atcggtagac ctcggccgtc gcggcgcttg
     2281 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     2341 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacagct ttgtttgtac
     2401 tatcaacagg ttgaactgct gatcaacaga tcctctacgc ggccgcggta ccataacttc
     2461 gtatagcata cattatacga agttatctgg ttttactagt atcgattcgc gacctactcc
     2521 ggaatattaa tagatcactc gagggatcca agctggccgc acgtctccct atcagtgata
     2581 gagaagtcga cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct
     2641 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     2701 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     2761 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     2821 gagctcccta tcagtgatag agaaggtacc ccctatataa gcagagctcg tttagtgaac
     2881 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     2941 cgatccagcc tccgcggtgg cggccatcac aagtcatacc gtcccaccat cgggcgcgga
     3001 tcccggtccg aagcgcgcgg aattcaaagg atgccagagc cagcgaagtc tgctcccgcc
     3061 ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc
     3121 aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac
     3181 cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt
     3241 ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc
     3301 acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac
     3361 gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagga tccaccggtc
     3421 gccaccatgg ccgaagggag cgtggcaaga cagcccgatc tgctgacatg cgacgacgaa
     3481 ccaatccaca ttccaggagc tatccagcct catggactgc tgctggctct ggccgctgac
     3541 atgactatcg tggcaggctc tgataacctg ccagagctga ccgggctggc tatcggagca
     3601 ctgattggcc ggtccgcagc cgacgtgttc gattctgaga ctcacaatag actgaccatc
     3661 gccctggctg aacctggggc tgcagtggga gccccaatta ccgtcggctt cacaatgcga
     3721 aaggacgctg gctttatcgg gtcctggcac cgccatgatc agctgatttt tctggagctg
     3781 gaaccacctc agagagacgt ggcagagcca caggctttct ttcggagaac aaacagcgcc
     3841 atcaggcgcc tgcaggccgc tgagactctg gaatccgctt gcgcagccgc tgcacaggaa
     3901 gtgaggaaga tcaccgggtt cgatcgcgtc atgatctaca gattcgcctc agactttagc
     3961 ggagaagtga tcgcagaaga tcgctgtgcc gaggtcgaaa gtaaactggg cctgcattac
     4021 ccagcctcaa ccgtgcctgc acaggcacga cggctgtata caatcaaccc cgtgcggatc
     4081 attcccgaca ttaattacag accagtgccc gtcacacctg acctgaaccc agtgactggg
     4141 aggcccatcg atctgagctt cgccattctg cgcagcgtgt cccctgtcca cctggagttt
     4201 atgaggaata tcggaatgca tggcaccatg tctatcagta ttctgagagg cgaaaggctg
     4261 tgggggctga tcgtgtgcca ccatcggaca ccttactatg tggacctgga cggcaggcag
     4321 gcttgtgaac tggtggctca ggtgctggct tggcagattg gcgtgatgaa ggaggaataa
     4381 cctacgtcga cgagctcact agtcgcggcc gctttcgaat ctagagcctg cagtctcgac
     4441 aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta
     4501 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4561 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4621 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4681 aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctagaagatc cggctgctaa
     4741 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tatcataacc
     4801 cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt catttttaat
     4861 tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact cttccctaaa
     4921 taatccttaa aaactccatt tccacccctc ccagttccca actattttgt ccgcccacag
     4981 cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc catgtgacaa
     5041 accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg tcatctcttc
     5101 g
//
