LOCUS       sequence-273992-        4512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor template to insert 5xTet operator sequence into downstream of 
            HoxC cluster..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4512)
  AUTHORS   Oksuz O, Narendra V, Lee CH, Descostes N, LeRoy G, Raviram R, 
            Blumenberg L, Karch K, Rocha PP, Garcia BA, Skok JA, Reinberg D
  TITLE     Capturing the Onset of PRC2-Mediated Repressive Domain Formation.
  JOURNAL   Mol Cell. 2018 Jun 21;70(6):1149-1162.e5. doi: 
            10.1016/j.molcel.2018.05.023.
  PUBMED    29932905
REFERENCE   2  (bases 1 to 4512)
  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"; doi: 
            "10.1016/j.molcel.2018.05"; journalName: "Mol Cell"; date: 
            "2018-06-21- 21"; volume: "70"; issue: "6"; pages: "1149-1162.e5"
COMMENT     Sequence Label: HoxC-downstream-5xTetO Donor
FEATURES             Location/Qualifiers
     source          1..4512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    521..542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(557..562,563..580,581..587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    595..611
                     /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)."
     primer_bind     600..622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     619..635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     619..635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1163..1181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1199..1217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1235..1253
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1271..1289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1307..1325
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1814..1833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1815..1833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1840..1857)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1840..1856)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1849..1871)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1994..2296
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2175..2194
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2645..3439
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2645..3439
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2699..2718)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3309..3328
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3639..3648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3645..4019
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      join(4157..4512,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      541 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      601 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctatttaggt
      661 gacgcgttag aatactcaag ctatgcatca agcttggtac cgagctcgga tccactagta
      721 acggccgcca gtgtgctgga attcaggtgt ggtggcacac acttgccatc tcagcaatgg
      781 gaacacggag agagaaggat gctatggcta ggtcactggc aacctccgac cagtgagatc
      841 ctgtctcaaa gagataaagc cactagaagc tgacctctca cctccatatg cagatacact
      901 cacgtgcacc atgcaccaac agatatgcac ctgcacatgt gcgaatgcaa acgcatgcac
      961 agagggtgga cctacctggt tgagataggt cacaagggaa agatattgga tgtttatacc
     1021 cagccaccgg ttcctctttg cttcctgctc tgctgtgaca tgaaggacat ctaccaagcc
     1081 cctcctccca cccctcccac tgtgaactaa gctgattcac caggctttcc tcaccatgat
     1141 ggattagacc acgagtcaaa attccctatc agtgatagag aagtttaggc ggactgtttc
     1201 cctatcagtg atagagaacg tatctgagtt ttactcccta tcagtgatag agattcctga
     1261 aaatgctcca tccctatcag tgatagagat tgatcatact ggacgttccc tatcagtgat
     1321 agagatgaga cctggttgcc caggtgctta ggttagtcct tatcctttct ccttcacccc
     1381 aaagataggt aactgcaaat agtgaatcct gggtggggta gtcatcactg agctgtatca
     1441 gttagcagtg attagcagtt atcatatctg cctcaccata gcctctacca ccccagagaa
     1501 cagacacagg ttgggtacca gctcttcact cacccttccc cagtctccca ggattctcac
     1561 ttaggagata acaggagctc agaagcagca agcctcgtgg aggtcccata tcccccattt
     1621 cagtctgctt ggtcctcaag ctctggacat ggagaattgc tgaatacatg ctttttgacc
     1681 cttaagacct tggtgcccct gggggaagtc agcaggagct ttaccatctt ttcaatcctg
     1741 gggtctgcct gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag
     1801 agggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     1861 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1921 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1981 cctatacgta cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     2041 tgtggatgta cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc
     2101 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     2161 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     2221 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     2281 gttctgggga atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct
     2341 tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     2401 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     2461 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     2521 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     2581 gccaaggatc tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt
     2641 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2701 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2761 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2821 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2881 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2941 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3001 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3061 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3121 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3181 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3241 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3301 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3361 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3421 tcttgacgag ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct
     3481 tacgcatctg tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg
     3541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3601 aaccctgata aatgcttcaa taatagcacg tgaggagggc caccatggcc aagttgacca
     3661 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     3721 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     3781 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     3841 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     3901 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     3961 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     4021 acgtgctaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4501 gcttggagcg aa
//
