pQZ8

LOCUS       pQZ8            6129 bp DNA     circular SYN 11-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pQZ008
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..61
                     /label=8x F
     enhancer        259..638
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..842
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        887..905
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(935..955)
                     /label=8x R
     protein_bind    1050..1245
                     /label=4x C/D-Box
                     /bound_moiety="L7Ae"
     primer_bind     1254..1285
                     /label=DDSEAP010 F
     primer_bind     complement(1264..1283)
                     /label=8x R
     primer_bind     1266..1285
                     /label=DDSEAP3.7 F
     CDS             1286..2788
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     primer_bind     complement(2827..2846)
                     /label=DDSEAP R
                     /note="同源重组，保留EcoRⅠ"
     polyA_signal    2846..2967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3632..4220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4391..5251)
                     /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"
     promoter        complement(5252..5356)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5383..5838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5840..5908)
                     /label=LacZ alpha
     polyA_signal    5969..6017
                     /note="synthetic polyadenylation signal"
     misc_feature    6031..6122
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tcccgatccc ctatggtgca
       61 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      121 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      181 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      241 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      301 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      361 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      421 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      481 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      541 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      601 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      661 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      721 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      781 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      841 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      901 atagggagac ccaagctggc tagcgtttaa acttaagctt caacaactac tagtttaacg
      961 taacatcgac agaccagtgg aagtaatact gcacgctagc taatagcaac cacacaaaca
     1021 atcaaagttt aaacgacaac gaaacttagg caacaaacaa atcaccaggg gcgtgatccg
     1081 aaaggtgacc ctggtgaaag ccttagccta ggggcgtgat ccgaaaggtg accctaggca
     1141 caagaaattc tacggggcgt gatccgaaag gtgacccgta gaatcacaaa gactacaggg
     1201 ggcgtgatcc gaaaggtgac ccctgtagac tcaaacttaa aaaggatatc ttcagcaaac
     1261 ttgaaccggt aagaattcgc ccaccatgct gctgctgctg ctgctgctgg gcctgaggct
     1321 acagctctcc ctgggcatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     1381 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     1441 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     1501 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     1561 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     1621 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     1681 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     1741 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     1801 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     1861 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     1921 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     1981 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2041 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2101 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     2161 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     2221 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     2281 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     2341 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     2401 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     2461 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     2521 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     2581 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     2641 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     2701 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     2761 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttactcta gagtcggggc
     2821 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     2881 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2941 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3001 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     3061 atcgataagg atctgaacga tggagcggag aatgggcgga actgggcgga gttaggggcg
     3121 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     3181 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     3241 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     3301 acacattcca cagcggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     3361 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     3421 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     3481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4201 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4381 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4681 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5041 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5221 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5341 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     5401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5821 tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     5881 cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta
     5941 aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg
     6001 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     6061 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     6121 ctatcgata
//