LOCUS       sequence-212155-        3793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inserts a polyA site to end transcription of a targetted gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3793)
  AUTHORS   Engreitz JM, Haines JE, Perez EM, Munson G, Chen J, Kane M, McDonel 
            PE, Guttman M, Lander ES
  TITLE     Local regulation of gene expression by lncRNA promoters, 
            transcription and splicing.
  JOURNAL   Nature. 2016 Nov 17;539(7629):452-455. doi: 10.1038/nature20149. 
            Epub 2016 Oct 26.
  PUBMED    27783602
REFERENCE   2  (bases 1 to 3793)
  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: 
            "10.1038/nature20149"; journalName: "Nature"; date: "2016-11-17- 
            17"; volume: "539"; issue: "7629"; pages: "452-455"
COMMENT     Sequence Label: pstop
FEATURES             Location/Qualifiers
     source          1..3793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    866..1000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1120..1254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1374..1508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1605..1623)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1644..1660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1644..1660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1657..1679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1668..1684
                     /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(1692..1698,1699..1716,1717..1722))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1737..1758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1875..1892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2046..2634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2126..2145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2805..3596,3597..3665))
                     /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     3428..3447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3666..3770)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 gggctgcagg aattcgatat tgaaccggtg cctagagaat cgcgatgaat aaatgaaagc
      721 ttgcagatct gcgactctag aggatctgcg actctagagg atcataatca gccataccac
      781 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
      841 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
      901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      961 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgcgactcta gaggatcata
     1021 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1081 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgcgac
     1261 tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1321 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1381 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1441 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1501 tctggatccc catcaagctg atccggaacc cttaatcgcc ctcgatctcg aactatcaag
     1561 cttatcgata ccgtcgaggg ggggcccggt accagctttt gttcccttta gtgagggtta
     1621 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1681 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1741 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1801 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1861 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1921 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1981 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2041 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2101 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2161 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2221 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2281 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2341 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2401 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2461 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2521 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2581 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2641 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2701 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2761 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2821 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2881 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2941 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3001 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3061 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3121 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3181 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3241 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3301 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3361 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3421 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3481 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3541 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3601 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3661 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3721 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3781 cgaaaagtgc cac
//
