LOCUS       sequence-487913-        4222 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Fragmid fragment: (CROPseq) cloning with mod1 LTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4222)
  TITLE     Fragmid
REFERENCE   2  (bases 1 to 4222)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AA628
FEATURES             Location/Qualifiers
     source          1..4222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    23..611
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(76..96)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(494..505)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        767..1015
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     767..787
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     938..957
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     LTR             1175..1355
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1381..1398)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1387..1611
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1711..1727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1711..1727)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1724..1746)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1735..1751
                     /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(1759..1765,1766..1783,1784..1789))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1942..1959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2113..2701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2193..2212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2879..3688)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3602..3621
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3680..3699
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3830..3852)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3952..3971
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4165..4187
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
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 cgcttttttc gtctcagttt cagagctatg ctggaaacag catagcaagt tgaaataagg
     1081 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttaa gcttggcgta
     1141 actagatctt gagacactgc tttttgcttg tactgggtct ctctggttag accagatctg
     1201 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     1261 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     1321 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     1381 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1441 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1501 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     1561 aggattggga agataatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     1621 cggaaagaac cagcgtccag tcttcggaac caaatcggat cccgggcccg tcgactgcag
     1681 aggcctgcat gcaagcttgg tgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1741 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1801 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1861 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1921 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2281 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2521 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2821 atgaagtttt aaatcaagcc caatctgaat aatgttacaa ccaattaacc aattctgatt
     2881 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     2941 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     3001 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     3061 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     3121 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     3181 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     3241 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     3301 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     3361 cttctaatac ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat
     3421 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     3481 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     3541 actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat
     3601 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     3661 tcgacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     3721 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     3781 gagacacggg ccagagctgc atcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3841 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3901 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3961 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4021 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4081 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     4141 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagagaat
     4201 tcgagctcgg tacctcgcga at
//
