LOCUS       sequence-200632-        3195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Standard for qPCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3195)
  AUTHORS   Yoder KE, Fishel R
  TITLE     Real-time quantitative PCR and fast QPCR have similar sensitivity 
            and accuracy with HIV cDNA late reverse transcripts and 2-LTR 
            circles.
  JOURNAL   J Virol Methods. 2008 Nov;153(2):253-6. doi: 
            10.1016/j.jviromet.2008.07.032. Epub 2008 Sep 17.
  PUBMED    18762215
REFERENCE   2  (bases 1 to 3195)
  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: 
            "10.1016/j.jviromet.2008.07.032"; journalName: "J Virol Methods"; 
            date: "2008-11"; volume: "153"; issue: "2"; pages: "253-6"
COMMENT     Sequence Label: pG-HIV-SIN-2LTR
FEATURES             Location/Qualifiers
     source          1..3195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    521..542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(557..562,563..580,581..587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    595..611
                     /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)."
     primer_bind     600..622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     619..635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     619..635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        653..671
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     653..670
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      945..954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      949..958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(988..1007)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(989..1007)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1014..1031)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1014..1030)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1023..1045)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1171..1626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1258..1277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1468..1489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1704..1808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1809..1877,1878..2669)
                     /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     complement(2027..2046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2840..3195,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      541 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      601 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctatttaggt
      661 gacactatag aatactcaag ctatgcatcc aacgcgttgg gagctctccc atatggtcga
      721 cctgcaggcg gccgcgaatt cactagtgat tagagagacc cagtacaagc aaaaagcaga
      781 tcttgtcttc gttgggagtg aattagccct tccagttgac taaaagggtc tgagggatct
      841 ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct
      901 ttattgaggc ttaagcagtg ggttccctaa atcgaattcc cgcggccgcc atggcggccg
      961 ggagcatgcg acgtcgggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     1021 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1081 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1141 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     1201 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1261 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1321 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1381 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1441 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1501 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1561 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     1621 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag
     1681 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1741 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1801 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1861 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1921 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1981 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2041 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2101 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2161 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2221 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2281 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2341 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2401 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2461 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2521 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2581 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2641 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2701 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2761 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2821 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2881 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2941 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3001 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3061 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3121 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3181 ccagcttgga gcgaa
//
