LOCUS       sequence-177430-        7531 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 7531)
  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 7531)
  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..7531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     complement(1810..1827)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1816..2040
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2125..2242
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2288..2321
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2544..2563)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2545..2563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2573..2590)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2573..2589)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2582..2604)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2731..3186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2818..2837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3028..3049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3212..3316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3317..3385,3386..4177)
                     /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     complement(3535..3554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4348..4936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4837..4856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5090..5107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5224..5245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5260..5265,5266..5283,5284..5290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5298..5314
                     /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     5303..5325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5322..5338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5322..5338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5357..5377
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5359..5377
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   5400..5434
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             complement(5917..6942)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(5917..6942)
                     /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(6961..7460)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     7402..7425
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    7483..7516
                     /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 gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cctcgacggt atcgataagc
     1741 tatcacaagt ttgtacattc gaaggatcca cgcgtctcga gttaacggta cctctagaga
     1801 attcgctagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1861 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1921 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1981 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2041 cttctgaggc ggttaattaa aataaaacta gagcgggccc ggtactaacc aaactggatc
     2101 tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc
     2161 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc
     2221 aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctgc agagcggccg
     2281 cgtcgagata acttcgtata atgtatgcta tacgaagtta tgttttgtta ctttatagaa
     2341 gaaattttga gtttttgttt ttttttaata aataaataaa cataaataaa ttgtttgttg
     2401 aatttattat tagtatgtaa gtgtaaatat aataaaactt aatatctatt caaattaata
     2461 aataaacgtc aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc
     2521 tagggttaaa gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     2581 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2641 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2701 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     2761 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2821 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2881 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2941 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3001 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3061 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3121 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3181 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3241 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3301 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3361 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3421 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3481 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3541 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3601 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3661 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3721 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3781 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3841 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3901 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3961 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4021 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4081 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4141 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4201 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4261 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4981 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5041 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5101 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5161 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5221 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5281 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5341 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg gtaccttaac
     5401 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
     5461 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
     5521 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
     5581 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
     5641 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
     5701 tagatatcca actatgaaac attatcataa agcaatagta aagagccttc agtaaagagc
     5761 aggcatttat ctaatcccac cccaccccca cccccgtagc tccaatcctt ccattcaaaa
     5821 tgtaggtact ctgttctcac ccttcttaac aaagtatgac aggaaaaaac ttccatgtgg
     5881 acatctttat tgtttaatag atcatcaatt tctcgactat tcctttgccc tcggacgagt
     5941 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     6001 cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     6061 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     6121 gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc
     6181 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     6241 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     6301 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     6361 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     6421 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     6481 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     6541 gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac
     6601 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     6661 acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg
     6721 aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     6781 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     6841 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     6901 aaacttctcg acagacgtcg cggtgagttc aggctttttc atggtggcgg ctggatcggt
     6961 cgaaaggccc ggagatgagg aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt
     7021 gcgagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc
     7081 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa
     7141 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca
     7201 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct
     7261 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac
     7321 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc
     7381 cacttgtgta gcgccaagtg ccagcggggc tgctaaagcg catgctccag actgccttgg
     7441 gaaaagcgcc tcccctaccc ggtagaattc atcgatgata tcataacttc gtataatgta
     7501 tgctatacga agttatgcgg ccctcgaggt c
//
