LOCUS       sequence-393237-        4644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Holding vector containing the contiguous 5'UTR and ORF1 regions of 
            human L1 (L1.3) to facilitate subcloning of ORF1 mutants into the 
            full-length L1 reporter pRTC2-puro (Cook et al., 2015)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4644)
  AUTHORS   Cook PR, Jones CE, Furano AV
  TITLE     Phosphorylation of ORF1p is required for L1 retrotransposition.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4298-303. doi: 
            10.1073/pnas.1416869112. Epub 2015 Mar 23.
  PUBMED    25831499
REFERENCE   2  (bases 1 to 4644)
  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.1416869112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-04-7- 7"; volume: "112"; issue: "14"; pages: "4298-303"
COMMENT     Sequence Label: p5'UTR-ORF1
FEATURES             Location/Qualifiers
     source          1..4644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2031..2053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2153..2172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2366..2388
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2380..2397
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2381..2397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4425..4441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4425..4441)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4438..4460)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4449..4465
                     /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(4473..4479,4480..4497,4498..4503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4518..4539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2161 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2221 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2281 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2341 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     2401 aagcttgcat gcggatccat gcgtacgggg aggagccaag atggccgaat aggaacagct
     2461 ccggtctaca gctcccagcg tgagcgacgc agaagacggt gatttctgca tttccatctg
     2521 aggtaccggg ttcatctcac tagggagtgc cagacagtgg gcgcaggcca gtgtgtgtgc
     2581 gcaccgtgcg cgagccgaag cagggcgagg cattgcctca cctgggaagc gcaaggggtc
     2641 agggagttcc ctttctgagt caaagaaagg ggtgacggtc gcacctggaa aatcgggtca
     2701 ctcccacccg aatattgcgc ttttcagacc ggcttaagaa acggcgcacc acgagactat
     2761 atcccacacc tggctcggag ggtcctacgc ccacggaatc tcgctgattg ctagcacagc
     2821 agtctgagat caaactgcaa ggcggcaacg aggctggggg aggggcgccc gccattgccc
     2881 aggcttgctt aggtaaacaa agcagccggg aagctcgaac tgggtggagc ccaccacagc
     2941 tcaaggaggc ctgcctgcct ctgtaggctc cacctctggg ggcagggcac agacaaacaa
     3001 aaagacagca gtaacctctg cagacttaag tgtccctgtc tgacagcttt gaagagagca
     3061 gtggttctcc cagcacgcag ctggagatct gagaacgggc agacagactg cctcctcaag
     3121 tgggtccctg actcctgacc cccgagcagc ctaactggga ggcacccccc agcaggggca
     3181 cactgacacc tcacacggca gggtattcca acagacctgc agctgagggt cctgtctgtt
     3241 agaaggaaaa ctaacaacca gaaaggacat ctacaccgaa aacccatctg tacatcacca
     3301 tcatcaaaga ccaaaagtag atgcggccgc aaagatgggg aaaaaacaga acagaaaaac
     3361 tggaaactct aaaacgcaga gcgcctctcc tcctccaaag gaacgcagtt cctcaccagc
     3421 aacggaacaa agctggatgg agaatgattt tgacgagctg agagaagaag gcttcagacg
     3481 atcaaattac tctgagctac gggaggacat tcaaaccaaa ggcaaagaag ttgaaaactt
     3541 tgaaaaaaat ttagaagaat gtataactag aataaccaat acagagaagt gcttaaagga
     3601 gctgatggag ctgaaaacca aggctcgaga actacgtgaa gaatgcagaa gcctcaggag
     3661 ccgatgcgat caactggaag aaagggtatc agcaatggaa gatgaaatga atgaaatgaa
     3721 gcgagaaggg aagtttagag aaaaaagaat aaaaagaaat gagcaaagcc tccaagaaat
     3781 atgggactat gtgaaaagac caaatctacg tctgattggt gtacctgaaa gtgatgtgga
     3841 gaatggaacc aagttggaaa acactctgca ggatattatc caggagaact tccccaatct
     3901 agcaaggcag gccaacgttc agattcagga aatacagaga acgccacaaa gatactcctc
     3961 gagaagagca actccaagac acataattgt cagattcacc aaagttgaaa tgaaggaaaa
     4021 aatgttaagg gcagccagag agaaaggtcg ggttaccctc aaaggaaagc ccatcagact
     4081 aacagtggat ctctcggcag aaaccctaca agccagaaga gagtgggggc caatattcaa
     4141 cattcttaaa gaaaagaatt ttcaacccag aatttcatat ccagccaaac taagcttcat
     4201 aagtgaagga gaaataaaat actttataga caagcaaatg ttgagagatt ttgtcaccac
     4261 caggcctgcc ctaaaagagc tcctgaagga agcgctaaac atggaaagga acaaccggta
     4321 ccagccgctg caaaatcatg ccaaaatgta aagaccatcg agactaggaa gaaactgcat
     4381 caactaatga gcaaaatccc gggacgcgtg aattcgtaat catggtcata gctgtttcct
     4441 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4501 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4561 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4621 agaggcggtt tgcgtattgg gcgc
//
