LOCUS       sequence-6939-ex        5331 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 5331)
  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 5331)
  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 2P
FEATURES             Location/Qualifiers
     source          1..5331
                     /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    2048..2169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2059..2078)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2113..2132
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2491..2511
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2663..2680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2834..3422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2914..2933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3593..4384,4385..4453))
                     /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     4216..4235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4454..4558)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4585..5040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4672..4691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4882..4903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5171..5219
                     /note="synthetic polyadenylation signal"
     misc_feature    5233..5324
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5273..5292
                     /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 tcgcgggccc tagagtcggg gcggccggcc gcttcgagca
     2041 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2101 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2161 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2221 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatctgaac
     2281 gatggagcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     2341 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     2401 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     2461 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagcggat
     2521 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     2581 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     2641 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4561 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4621 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4681 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4741 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4801 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4861 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4921 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4981 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5041 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5101 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
     5161 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     5221 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     5281 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat a
//
