LOCUS       sequence-448490-        3310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3310)
  AUTHORS   Chen B, Cepko CL
  TITLE     HDAC4 regulates neuronal survival in normal and diseased retinas.
  JOURNAL   Science. 2009 Jan 9. 323(5911):256-9.
  PUBMED    19131628
REFERENCE   2  (bases 1 to 3310)
  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: "Science. 
            2009 Jan 9. 323(5911):256-9."
COMMENT     Sequence Label: PBS/pU6-HIF1 alpha RNAi plasmid 2
FEATURES             Location/Qualifiers
     source          1..3310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..25
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     17..34
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     18..34
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     44..63
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        44..62
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     80..96
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        93..406
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     325..345
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(487..503)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(540..560)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(540..558)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(579..595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(579..595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(592..614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    603..619
                     /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(627..633,634..651,652..657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    672..693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(810..827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(981..1569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1061..1080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1740..2531,2532..2600))
                     /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     2363..2382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2601..2705)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2731..3186)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2868..2889)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3080..3099
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
       61 gggcgaattg ggtacccgct ctagaactag tggatccgac gccgccatct ctaggcccgc
      121 gccggccccc tcgcacagac ttgtgggaga agctcggcta ctcccctgcc ccggttaatt
      181 tgcatataat atttcctagt aactatagag gcttaatgtg cgataaaaga cagataatct
      241 gttcttttta atactagcta cattttacat gataggcttg gatttctata agagatacaa
      301 atactaaatt attattttaa aaaacagcac aaaaggaaac tcaccctaac tgtaaagtaa
      361 ttgtgtgttt tgagactata aatatccctt ggagaaaagc cttgtttggg gttgaaactc
      421 aagcaactgg tgtgctgtcc cagttgcttg agtttcaacc cctttttgaa ttcctgcagc
      481 ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
      541 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
      601 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      661 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      721 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      781 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2701 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     2761 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2821 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2881 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2941 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3001 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3061 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3121 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3181 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3241 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3301 cgccagggtt
//
