LOCUS       sequence-205531-        3794 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 3794)
  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 3794)
  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..3794
                     /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    867..1001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1121..1255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1375..1509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1606..1626)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1606..1624)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1645..1661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1645..1661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1658..1680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1669..1685
                     /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(1693..1699,1700..1717,1718..1723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1738..1759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1876..1893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2047..2635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2127..2146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2806..3597,3598..3666))
                     /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     3429..3448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3667..3771)
                     /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 gccntaccac
      781 attttgtaga ggttttactn gctttaaaaa acctcccaca cctccccctg aacctgaaac
      841 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      961 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgcgactct agaggatcat
     1021 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1081 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1141 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1201 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgcga
     1261 ctctagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1321 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1381 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1441 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     1501 gtctggatcc ccatcaagct gatccggaac ccttaatcgc cctcgatctc gaactatcaa
     1561 gcttatcgat accgtcgagg gggggcccgg taccagcttt tgttcccttt agtgagggtt
     1621 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1681 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1741 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1801 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1861 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1921 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1981 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2041 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2101 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2161 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2221 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2281 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2341 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2401 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2461 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     2521 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2581 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2641 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2701 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2761 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2941 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3661 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3781 ccgaaaagtg ccac
//
