LOCUS       sequence-362679-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression empty backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  AUTHORS   Beucher A, Miguel-Escalada I, Balboa D, De Vas MG, Maestro MA, 
            Garcia-Hurtado J, Bernal A, Gonzalez-Franco R, Vargiu P, Heyn H, 
            Ravassard P, Ortega S, Ferrer J
  TITLE     The HASTER lncRNA promoter is a cis-acting transcriptional 
            stabilizer of HNF1A.
  JOURNAL   Nat Cell Biol. 2022 Oct 6. pii: 10.1038/s41556-022-00996-8. doi: 
            10.1038/s41556-022-00996-8.
  PUBMED    36202974
REFERENCE   2  (bases 1 to 5175)
  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: "Nat Cell 
            Biol. 2022 Oct 6. pii: 10.1038/s41556-022-00996-8. doi: 
            10.1038/s41556-022-00996-8."
COMMENT     Sequence Label: pcDNA3.1-flag-UTR
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    72..296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      342..770
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(429..448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     639..660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(779..799)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        784..1113
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      964..1099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1026..1045
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1161..1208
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1227..1601
                     /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"
     polyA_signal    1731..1852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1768..1787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1822..1841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1901..1917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1901..1917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1914..1936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1925..1941
                     /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(1949..1955,1956..1973,1974..1979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1994..2015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2132..2149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2303..2891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2383..2402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3062..3853,3854..3922))
                     /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     3685..3704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3923..4027)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4102..4121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4293..4672
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4673..4876
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4827..4847
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4921..4940
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4921..4939
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5023..5046
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     3'UTR           5050..5175
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
ORIGIN
        1 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
       61 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
      121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
      181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
      241 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
      301 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
      361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      541 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      661 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      721 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
      781 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      841 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      901 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      961 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1021 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     1081 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     1141 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     1201 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     1261 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     1321 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     1381 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     1441 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     1501 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     1561 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     1621 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     1681 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1741 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1801 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1861 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1921 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1981 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2041 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2101 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2161 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2221 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2281 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2701 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3001 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3061 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3121 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3181 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3241 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3301 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3361 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3421 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3481 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3541 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3601 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3661 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3721 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3781 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3841 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3901 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3961 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4021 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     4081 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     4141 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     4201 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     4261 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     4321 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4381 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4441 gacgtatgtt cccatagyaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4501 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4561 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4621 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     4681 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4741 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4801 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4861 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     4921 taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc
     4981 gagctcggat ccactagtcc agtgtggtgg gaattcacta gtgactacaa agacgatgac
     5041 gataaataaa accagcctca agaacacccg aatggagtct ctaagctaca taataccaac
     5101 ttacacttta caaaatgttg tcccccaaaa tgtagccatt cgtatctgct cctaataaaa
     5161 agaaagtttc ttcac
//
