LOCUS       sequence-6940-ex        5356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   Doench JG, Petersen CP, Sharp PA
  TITLE     siRNAs can function as miRNAs.
  JOURNAL   Genes Dev. 2003 Feb 15. 17(4):438-42.
  PUBMED    12600936
REFERENCE   2  (bases 1 to 5356)
  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"; journalName: "Genes Dev. 
            2003 Feb 15. 17(4):438-42."
COMMENT     Sequence Label: pGL3 CXCR4 3P
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      274..283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             280..1932
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(346..364)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2073..2194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2084..2103)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2138..2157
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2516..2536
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2688..2705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2859..3447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2939..2958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3618..4409,4410..4478))
                     /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     4241..4260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4479..4583)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4610..5065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4697..4716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4907..4928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5196..5244
                     /note="synthetic polyadenylation signal"
     misc_feature    5258..5349
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5298..5317
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
      301 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
      361 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
      421 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
      481 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
      541 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
      601 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
      661 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
      721 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
      781 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
      841 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
      901 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
      961 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     1021 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     1081 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     1141 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     1201 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     1261 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     1321 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     1381 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     1441 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     1501 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     1561 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     1621 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     1681 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     1741 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     1801 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     1861 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     1921 atcgccgtgt aattctagac tcgagccgga agttttcact ccagctaaca ccggaagttt
     1981 tcactccagc taacaccgga agttttcact ccagctaaca ccggatcgcg ggccctagag
     2041 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2101 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2161 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2221 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2281 tggtaaaatc gataaggatc tgaacgatgg agcggagaat gggcggaact gggcggagtt
     2341 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     2401 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     2461 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     2521 aactgacaca cattccacag cggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2581 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2641 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4621 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4681 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4741 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4801 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4861 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4921 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4981 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5041 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     5101 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     5161 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     5221 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     5281 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     5341 catttctcta tcgata
//
