LOCUS       sequence-394518-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For lncRNA overexpression (in trans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  AUTHORS   Yin Y, Yan P, Lu J, Song G, Zhu Y, Li Z, Zhao Y, Shen B, Huang X, 
            Zhu H, Orkin SH, Shen X
  TITLE     Opposing Roles for the lncRNA Haunt and Its Genomic Locus in 
            Regulating HOXA Gene Activation during Embryonic Stem Cell 
            Differentiation.
  JOURNAL   Cell Stem Cell. 2015 May 7;16(5):504-16. doi: 
            10.1016/j.stem.2015.03.007. Epub 2015 Apr 16.
  PUBMED    25891907
REFERENCE   2  (bases 1 to 7513)
  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.stem.2015.03.007"; journalName: "Cell Stem Cell"; date: 
            "2015-05-7- 7"; volume: "16"; issue: "5"; pages: "504-16"
COMMENT     Sequence Label: PB-CAG-BGHpA
FEATURES             Location/Qualifiers
     source          1..7513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1548..1570
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1632..1651
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1767..1784)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1773..1997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2082..2199
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2245..2278
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   2449..2511
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(2535..2553)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2574..2590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2574..2590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2587..2609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2598..2614
                     /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(2622..2628,2629..2646,2647..2652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2667..2688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2805..2822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2976..3564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3056..3075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3735..4526,4527..4595))
                     /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     4358..4377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4596..4700)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4726..5181)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4863..4884)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5075..5094
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5308..5330
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5322..5339
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5323..5339
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5346..5365
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5346..5364
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5383..5417
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             complement(5899..6924)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(5899..6924)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6943..7442)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6991..7010)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7384..7407
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    7465..7498
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gncgcgcgcc aggcggggcg gggcssggcg wskkgcksgs
      541 cbkkkygysb msstcrckcs kckcgcctgt gtgcgggagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg cgckgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
      961 cgggcggcbc scsgggcttt gtgcgctccg crgtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg gggctgsgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggccaggtgg gggtgccggg
     1201 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
     1261 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     1321 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
     1381 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     1441 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1501 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1561 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1621 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct
     1681 cgacggtatc gataagctat cacaagtttg tacattcgaa ggatccacgc gtctcgagtt
     1741 aacggtacct ctagagaatt cgctagcctc gactgtgcct tctagttgcc agccatctgt
     1801 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1861 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1921 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1981 tgcggtgggc tctatggctt ctgaggcggt taattaaaat aaaactagag cgggcccggt
     2041 actaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta
     2101 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     2161 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
     2221 tgtctgcaga gcggccgcgt cgagataact tcgtataatg tatgctatac gaagttatgt
     2281 tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat
     2341 aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat
     2401 atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt
     2461 ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaaagagc
     2521 tccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     2581 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2641 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2701 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2761 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2881 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2941 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3001 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3061 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3361 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3721 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3781 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3841 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3901 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3961 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4021 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4081 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4141 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4201 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4261 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4321 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4381 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4441 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4501 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4561 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     4741 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4801 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4861 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4921 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4981 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5041 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5101 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5161 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     5221 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5281 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5341 aattgtaata cgactcacta tagggcgaat tgggtacctt aaccctagaa agatagtctg
     5401 cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata gcgcgaatcc
     5461 gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg cgtgcttgtc
     5521 aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa atgacgcatg
     5581 attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg ttatttcatg
     5641 ttctacttac gtgataactt attatatata tattttcttg ttatagatat ccaactatga
     5701 aacattatca taaagcaata gtaaagagcc ttcagtaaag agcaggcatt tatctaatcc
     5761 caccccaccc ccacccccgt agctccaatc cttccattca aaatgtaggt actctgttct
     5821 cacccttctt aacaaagtat gacaggaaaa acttccatgt ggacatcttt attgtttaat
     5881 agatcatcaa tttctcgact attcctttgc cctcggacga gtgctggggc gtcggtttcc
     5941 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
     6001 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
     6061 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
     6121 gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa
     6181 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
     6241 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
     6301 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
     6361 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
     6421 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
     6481 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
     6541 gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
     6601 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
     6661 ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
     6721 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
     6781 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
     6841 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt
     6901 cgcggtgagt tcaggctttt tcatggtggc ggctggatcg gtcgaaaggc ccggagatga
     6961 ggaagaggag aacagcgcgg cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc
     7021 ggaggacctt cgggcgcccg ccccgcccct gagcccgccc ctgagcccgc ccccggaccc
     7081 accccttccc agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc
     7141 ccaaaggcct acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag
     7201 acgtgctact tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt
     7261 cctgactagg ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg
     7321 gcgcctaccg gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt
     7381 gcccagcggg gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac
     7441 ccggtagaat tcatcgatga tatcataact tcgtataatg tatgctatac gaagttatgc
     7501 ggccctcgag gtc
//
