LOCUS       sequence-227086-        5174 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 5174)
  AUTHORS   Krupnik VE, Sharp JD, Jiang C, Robison K, Chickering TW, Amaravadi 
            L, Brown DE, Guyot D, Mays G, Leiby K, Chang B, Duong T, Goodearl 
            AD, Gearing DP, Sokol SY, McCarthy SA
  TITLE     Functional and structural diversity of the human Dickkopf gene 
            family.
  JOURNAL   Gene. 1999 Oct 1. 238(2):301-13.
  PUBMED    10570958
REFERENCE   2  (bases 1 to 5174)
  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: "Gene. 1999 
            Oct 1. 238(2):301-13."
COMMENT     Sequence Label: pCS2-hDKK3-flag
FEATURES             Location/Qualifiers
     source          1..5174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1122..1145
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1192..1326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1216..1235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1270..1289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1439..1459)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1439..1457)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1478..1494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1478..1494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1491..1513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1502..1518
                     /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)."
     promoter        complement(join(1526..1532,1533..1550,1551..1556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1571..1592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1709..1726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1880..2468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1960..1979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2639..3430,3431..3499))
                     /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     3262..3281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3500..3604)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3630..4085)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3767..3788)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3979..3998
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4156..5140
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 atttaggtga cactatagaa tacaagctac ttgttctttt tgcaggatcc catcgattcg
       61 aattcgccac catgcagcgg cttggggcca ccctgctgtg cctgctgctg gcggcggcgg
      121 tccccacggc ccccgcgccc gctccgacgg cgacctcggc tccagtcaag cccggcccgg
      181 ctctcagcta cccgcaggag gaggccaccc tcaatgagat gttccgcgag gttgaggaac
      241 tgatggagga cacgcagcac aaattgcgca gcgcggtgga agagatggag gcagaagaag
      301 ctgctgctaa agcatcatca gaagtgaacc tggcaaactt acctcccagc tatcacaatg
      361 agaccaacac agacacgaag gttggaaata ataccatcca tgtgcaccga gaaattcaca
      421 agataaccaa caaccagact ggacaaatgg tcttttcaga gacagttatc acatctgtgg
      481 gagacgaaga aggcagaagg agccacgagt gcatcatcga cgaggactgt gggcccagca
      541 tgtactgcca gtttgccagc ttccagtaca cctgccagcc atgccggggc cagaggatgc
      601 tctgcacccg ggacagtgag tgctgtggag accagctgtg tgtctggggt cactgcacca
      661 aaatggccac caggggcagc aatgggacca tctgtgacaa ccagagggac tgccagccgg
      721 ggctgtgctg tgccttccag agaggcctgc tgttccctgt gtgcacaccc ctgcccgtgg
      781 agggcgagct ttgccatgac cccgccagcc ggcttctgga cctcatcacc tgggagctag
      841 agcctgatgg agccttggac cgatgccctt gtgccagtgg cctcctctgc cagccccaca
      901 gccacagcct ggtgtatgtg tgcaagccga ccttcgtggg gagccgtgac caagatgggg
      961 agatcctgct gcccagagag gtccccgatg agtatgaagt tggcagcttc atggaggagg
     1021 tgcgccagga gctggaggac ctggagagga gcctgactga agagatggcg ctgggggagc
     1081 ctgcggctgc cgccgctgca ctgctgggaa gggaagagat tgactacaag gacgacgacg
     1141 acaagtgagt cgacctcgag cctctagaac tatagtgagt cgtattacgt agatccagac
     1201 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1261 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1321 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1381 gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc
     1441 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1501 attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct
     1561 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1621 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1681 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1741 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1801 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2041 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2281 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2401 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2461 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2521 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2581 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2641 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2701 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2761 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2821 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2881 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2941 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3001 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3061 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3121 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3181 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3241 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3301 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3361 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3421 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3481 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3541 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3601 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     3661 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3721 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3781 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3841 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3901 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3961 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4021 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4081 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4141 ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc atgccaattc
     4201 aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc
     4261 aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc
     4321 aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt
     4381 ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata
     4441 atggcggcca tattggctat atgccaggat caatatatag gcaatatcca atatggccct
     4501 atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat
     4561 tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac
     4621 catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct
     4681 atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc
     4741 cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag
     4801 gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc
     4861 cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat
     4921 agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc agtactccca
     4981 ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca
     5041 atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc
     5101 ctaatgggag gtctatataa gcaatgctcg tttagggaac cgccattctg cctggggacg
     5161 tcggagcaag cttg
//
