LOCUS       pF_AA627                4211 bp    DNA     circular SYN 02-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4211)
  AUTHORS   jdoench
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    11..22
                     /label=GG-6F
     misc_feature    23..611
                     /label=WPRE_v1
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     misc_difference 561
                     /label=Silent
     misc_difference 616
                     /label=mod1 (G-->A)
                     /note="from Bock/Blainey/Shendure/512/722 vectors"
     misc_difference 619..675
                     /label=mod1 (+57nt)
                     /note="sequence from Bock/Blainey/Shendure/512/722 vectors"
     misc_difference 676..680
                     /label=mod1 (+5nt)
                     /note="sequence from Bock/Blainey/Shendure/512/722 vectors"
     misc_difference 712
                     /label=Silent
     promoter        767..1015
                     /label=U6_v1
     misc_feature    1012..1022
                     /label=BsmBI_v0
     misc_feature    1024..1029
                     /label=6T
     misc_feature    1046..1056
                     /label=BsmBI_v24
     misc_feature    1052..1111
                     /label=trRNA_v9 [Sp]
                     /note="Modified trRNA_v4 to include C/G from Blainey lab 
                     for primer compatibility for OPS readout. Short version of 
                     trRNA_v5."
     LTR             1164..1344
                     /label=5' LTR (truncated)
     polyA_signal    1376..1600
                     /label=bGHpolyA_v1
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1623..1634
                     /label=GG-9R
     primer_bind     complement(1700..1716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1724..1740
                     /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(1748..1754,1755..1772,1773..1778))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1793..1814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2102..2690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2868..3677)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 acatctagat gaagacggat tcaatcaacc tctggattac aaaatttgtg aaagattgac
       61 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
      121 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
      181 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
      241 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
      301 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
      361 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
      421 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
      481 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
      541 tctgcggcct cttccgcgtc ctcgccttcg ccctcagacg agtcggatct ccctttgggc
      601 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
      661 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatttc
      721 accattatcg tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc
      781 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
      841 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
      901 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
      961 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga
     1021 cgcttttttg tttcagagct agaaacgtct ctagcaagtt gaaataaggc tagtccgtta
     1081 tcaacttgaa aaagtggcac cgagtcggtg cttttttaag cttggcgtaa ctagatcttg
     1141 agacactgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     1201 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     1261 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     1321 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1381 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1441 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1501 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1561 gataatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     1621 agcgtccagt cttcggaacc aaatcggatc ccgggcccgt cgactgcaga ggcctgcatg
     1681 caagcttggt gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1741 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1801 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1861 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1921 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1981 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2041 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2281 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2461 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2521 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2581 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2821 aatcaagccc aatctgaata atgttacaac caattaacca attctgatta gaaaaactca
     2881 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     2941 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3001 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     3061 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     3121 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     3181 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     3241 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     3301 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3361 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3421 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     3481 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3541 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3601 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc
     3661 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3721 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacgggc
     3781 cagagctgca tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3841 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3901 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     3961 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4021 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     4081 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     4141 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt
     4201 acctcgcgaa t
//
