pSCM-N20-V3-ccdb

LOCUS       pSCM-N20-V3-ccdb               6995 bp DNA     circular SYN 23-JUN-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSCm-N20-V3-ccdb
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6995)
  AUTHORS   miao liu
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..6995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(682..1025)
                     /label=Promoter_P_1
                     /note="TEF promoter Ashbya gossypii TEF promoter"
     promoter        1032..1300
                     /label=Promoter_P_2
                     /note="SNR52 promoter promoter for the S. cerevisiae small 
                     nucleolar RNA  gene SNR52"
     primer_bind     1037..1056
                     /label=c-SCm-gRNA-F
     misc_feature    1311..2298
                     /label=ccdB
     primer_bind     2276..2296
                     /label=0227-SZgRNA-test-F
     misc_RNA        2309..2383
                     /label=RNA_1
                     /note="gRNA scaffold guide RNA scaffold for the 
                     Streptococcus pyogenes  CRISPR/Cas9 system"
     terminator      2388..2407
                     /label=Terminator_1
                     /note="SUP4 terminator"
     misc_feature    2408..2445
                     /label=Misc_Feature_2
                     /note="MCS"
     primer_bind     complement(2664..2683)
                     /label=0227-SZgRNA-test-R
     rep_origin      complement(2745..3333)
                     /direction=LEFT
                     /label=Rep_Origin_1
                     /note="ori high-copy-number ColE1/pMB1/pBR322/pUC origin of
                      replication"
     CDS             complement(3504..4364)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR confers resistance to ampicillin, 
                     carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5040..5064
                     /label=2u-test-F
     misc_feature    5041..5062
                     /label=Chro267-1-F
     rep_origin      5065..5942
                     /direction=RIGHT
                     /label=Rep_Origin_2
                     /note="2u ori yeast 2u plasmid origin of replication"
     primer_bind     complement(5923..5942)
                     /label=2u-test-R
     terminator      6445..6642
                     /label=Terminator_2
                     /note="TEF terminator Ashbya gossypii TEF terminator"
     CDS             complement(join(6648..6995,1..681))
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=aph(4)-Ia
                     /note="HygR confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
ORIGIN
        1 ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc
       61 cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc
      121 ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc
      181 catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca
      241 tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag
      301 atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag cgggcagttc
      361 ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag
      421 gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa
      481 gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac
      541 atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg
      601 catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt
      661 gagttcaggc tttttaccca tggttgttta tgttcggatg tgatgtgaga actgtatcct
      721 agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaatc ctaacctttt
      781 atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag gggagcgttt
      841 ccctgctcgc aggtccgcag cgaggagccg taatttttgc ttcgcgccgt gcggccatca
      901 aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta cgggcgacag
      961 tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc
     1021 atgtcactag ttctttgaaa agataatgta tgattatgct ttcactcata tttatacaga
     1081 aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact
     1141 ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac
     1201 aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt
     1261 tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc tgagacccag gtcgaccgac
     1321 agccttccaa atgttcttct caaacggaat cgtcgtatcc agcctactcg ctattgtcct
     1381 caatgccgta ttaaatcata aaaagaaata agaaaaagag gtgcgagcct cttttttgtg
     1441 tgacaaaata aaaacatcta cctattcata tacgctagtg tcatagtcct gaaaatcatc
     1501 tgcatcaaga acaatttcac aactcttata cttttctctt acaagtcgtt cggcttcatc
     1561 tggattttca gcctctatac ttactaaacg tgataaagtt tctgtaattt ctactgtatc
     1621 gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag aacatcaggt
     1681 taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct tccccgataa
     1741 cggagaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca tccccgatat
     1801 gcaccaccgg gtaaagttca cgggagactt tatctgacag cagacgtgca ctggccaggg
     1861 ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc acaaacagac
     1921 gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca gcccctgttc
     1981 tcgtcagcaa aagagccgtt catttcaata aaccgggcga cctcagccat cccttcctga
     2041 ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat ggttgtgctt
     2101 accagaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa
     2161 ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc gggtatacat
     2221 atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc
     2281 ttttagtaag ccggatccct gggtctcggt tttagagcta gaaatagcaa gttaaaataa
     2341 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtggtgcttt ttttgttttt
     2401 tatgtctaga tcctaggatc caagcttggt accgagctcg tcgacgaatt ctgcagatat
     2461 ccatcacact ggcggccgct cgagcatgca tctagagggc cgcaggccct gcattaatga
     2521 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3181 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgtgaccc acgctcaccg
     3661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4381 tgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
     4441 ttacttataa tacagttttt tgaaaagcta gcttatcgat gataagctgt caaacatgag
     4501 aattaattcc acggactata gactatacct agtatactcc gtctactgta cgatacactt
     4561 ccgctcaggt ccttgtcctt taacgaggcc ttaccactct tttgttactc tattgatcca
     4621 gctcagcaaa ggcagtgtga tctaagattc tatcttcgcg atgtagtaaa actagctaga
     4681 ccgagaaaga gactagaaat gcaaaaggca cttctacaat ggctgccatc attattatcc
     4741 gatgtgacgc tgcagcttct caatgatatt cgaatacgct ttgaggagat acagcctaat
     4801 atccgacaaa ctgttttaca gatttacgat cgtacttgtt acccatcatt gaattttgaa
     4861 catccgaacc tgggagtttt ccctgaaaca gatagtatat ttgaacctgt ataataatat
     4921 atagtctagc gctttacgga agacaatgta tgtatttcgg ttcctggaga aactattgca
     4981 tctattgcat aggtaatctt gcacgtcgca tccccggttc attttctgcg tttccatctt
     5041 gcacttcaat agcatatctt tgttaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     5101 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     5161 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     5221 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     5281 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     5341 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     5401 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     5461 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     5521 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     5581 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     5641 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     5701 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     5761 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     5821 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     5881 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     5941 atgggaagct ccaccccggt tgataatcag aaaagcccca aaaacaggaa gattgtataa
     6001 gcaaatattt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6061 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6121 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6181 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6241 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6301 aaagggagcc cccgatttag agcttgacgg ggaaagccct gatgaatatc ttgtggtagg
     6361 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag
     6421 tacagaagat taagtgagag atcccagtat agcgaccagc attcacatac gattgacgca
     6481 tgatattact ttctgcgcac ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa
     6541 aatataaatc acgctaacat ttgattaaaa tagaacaact acaatataaa aaaactatac
     6601 aaatgacaag ttcttgaaaa caagaatctt tttattgtca gtactgatta ttcctttgcc
     6661 ctcggacgag tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg
     6721 gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat
     6781 cggacgattg cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc
     6841 aagctctgat agagttggtc aagaccaatg cggagcatat acgcccggag ccgcggcgat
     6901 cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac
     6961 cacggcctcc agaagaagat gttggcgacc tcgta
//