LOCUS       sequence-158517-        4312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes partial human gamma- and beta-globing cDNA for use as QPCR 
            standard as published in PMID: 14592835...
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4312)
  AUTHORS   Bhanu NV, Trice TA, Lee YT, Miller JL
  TITLE     A signaling mechanism for growth-related expression of fetal 
            hemoglobin.
  JOURNAL   Blood. 2004 Mar 1;103(5):1929-33. Epub 2003 Oct 30.
  PUBMED    14592835
REFERENCE   2  (bases 1 to 4312)
  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: "Blood."; 
            date: "2004-03-1"; volume: "103(5)"; pages: "1929-33. Epub 2003 Oct 
            30"
COMMENT     Sequence Label: BGGLO QPCR PLASMID
FEATURES             Location/Qualifiers
     source          1..4312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(745..764)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(746..764)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(771..788)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(771..787)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(780..802)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      928..1356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1015..1034)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1225..1246
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1700..2494
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1754..1773)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2364..2383
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2512..2580,2581..3372)
                     /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(2730..2749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3543..4131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4032..4051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4285..4302
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 aatggattgc caaaacggtc accagcacat ttcccaggag cttgaagttc tcaggatcca
      301 catgcagctt gtcacagtgc agttcactca gctgggcaaa ggtgcccttg aggtcatcca
      361 ggtgctttat ggcatctccc aaggaagtca gcacctcctt gccatgtgcc ttgactttgg
      421 ggttgcccat gatggcagag gcagaggaca ggttgccact agtaacggcc gccagtgtgc
      481 tggaattcgc ccttctcatg gcaagaaagt gctcggtgcc tttagtgatg gcctggctca
      541 cctggacaac ctcaagggca cctttgccac actgagtgag ctgcactgtg acaagctgca
      601 cgtggatcct gagaacttca ggctcctggg caacgtgctg gtctgtgtgc tggcccatca
      661 ctttggcaaa gaattaaggg cgaattctgc agatatccat cacactggcg gccgctcgag
      721 catgcatcta gagggcccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
      781 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      841 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      901 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
      961 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1021 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1081 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1141 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1201 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1261 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1321 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaattcag ggcgcaaggg
     1381 ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat
     1441 gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt
     1501 agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga
     1561 accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
     1621 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac
     1681 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     1741 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     1801 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     1861 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     1921 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     1981 gggcgaagtg ccggggcagg atctcctgtc atcccacctt gctcctgccg agaaagtatc
     2041 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2101 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     2161 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     2221 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     2281 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     2341 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     2401 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     2461 cgccttctat cgccttcttg acgagttctt ctgaattgaa aaaggaagag tatgagtatt
     2521 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2581 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2641 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2701 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2761 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2821 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2881 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2941 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3001 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3061 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3121 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3181 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3241 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3301 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3361 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3421 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3481 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3661 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3721 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3781 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3841 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3901 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3961 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4021 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4081 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4141 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4201 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4261 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
