LOCUS       sequence-197848-        4623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pin nuclease domain with Pum cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4623)
  AUTHORS   Adamala KP, Martin-Alarcon DA, Boyden ES
  TITLE     Programmable RNA-binding protein composed of repeats of a single 
            modular unit.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2579-88. doi: 
            10.1073/pnas.1519368113. Epub 2016 Apr 26.
  PUBMED    27118836
REFERENCE   2  (bases 1 to 4623)
  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: 
            "10.1073/pnas.1519368113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-10- 10"; volume: "113"; issue: "19"; pages: 
            "E2579-88"
COMMENT     Sequence Label: [GG][Pin]
FEATURES             Location/Qualifiers
     source          1..4623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    710..831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(721..740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     775..794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1012..1467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1099..1118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1309..1330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1534..1553)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1653..1675
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1713..1731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1799..1903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1904..1972,1973..2764)
                     /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(2122..2141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2935..3523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3424..3443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3734..4113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4114..4317
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4268..4288
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4314..4338
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4453..4585
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4568..4587
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 agtacttaat acgactcact ataggctagc cgccaccatg cgagagagac cggatggcag
       61 agagtcgcta tctcatgtac cgatagtagc ggtctccacg cgagtggaca ccggaaacgg
      121 ctccatggaa ctcgagatca ggccactttt cctggtgcca gacactaatg gattcataga
      181 ccacctcgcc agcctcgcac gcctcttgga gtcccgcaag tatattctgg tagttccgct
      241 gatcgttatc aatgagttgg acggcttggc caagggacag gagactgacc atcgcgccgg
      301 tgggtacgcc cgcgtcgtgc aagagaaggc tagaaaatca atcgaattcc tggagcaacg
      361 cttcgaatca cgggattcct gcctgagggc tcttacctcc agaggcaacg agctggagtc
      421 tattgcattc cgcagcgagg acatcacagg gcagctgggc aataacgatg atcttattct
      481 gagctgttgt cttcactatt gtaaggacaa ggctaaagat ttcatgcccg ctagtaagga
      541 agaacccatt cggctgctgc gcgaagtcgt gctgctgaca gacgatcgca atctccgggt
      601 taaagccttg actcggaacg tgccagtgag ggatatcccc gccttcctga catgggctca
      661 ggtgggataa tctagagtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
      721 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
      781 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
      841 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
      901 taaaacctct acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa tagcgaagag
      961 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     1021 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1081 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1141 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1201 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1261 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1321 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1381 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1441 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     1501 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     1561 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     1621 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     1681 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     1741 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1801 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1861 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1921 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1981 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2041 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2101 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2161 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2221 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2281 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2341 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2401 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2461 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2521 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2581 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcgcgcggta tcattgcagc
     2641 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2701 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2761 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3061 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac agatcttcaa
     3601 tattggccat tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg
     3661 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
     3721 tgaccgccat gttggcattg attattgact agttattaat agtaatcaat tacggggtca
     3781 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3841 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     3901 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     3961 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
     4021 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
     4081 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat
     4141 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     4201 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
     4261 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     4321 ttagtgaacc gtcagatcac tagaagcttt attgcggtag tttatcacag ttaaattgct
     4381 aacgcagtca gtgcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga
     4441 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggaggcca atagaaactg
     4501 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     4561 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
     4621 tag
//
