LOCUS       Exported                6813 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    SOST/pcDNA3.1+
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6813)
  AUTHORS   Semenov M, Tamai K, He X
  TITLE     SOST is a ligand for LRP5/LRP6 and a Wnt signaling inhibitor.
  JOURNAL   J Biol Chem. 2005 Jul 22. 280(29):26770-5.
  PUBMED    15908424
REFERENCE   2  (bases 1 to 6813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6813
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        143..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"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /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
                     GLAPAELFARLKAGMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(3995..4014)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3996..4014)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4059..4262)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4088..4108)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4263..4642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4814..4833
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4908..5012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5013..5873
                     /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(5231..5250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6044..6632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6533..6552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6786..6803
                     /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 gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc ccgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgggtttaa acgggccctc tagactcgag cggccgccac
     2521 tgtgctggat atctgcagaa ttccaccaca ctggactagt ggatcttcat ttacccggag
     2581 acagggagag gctcttctgc gtgtagtggt tgtgcagagc ctcatgcatc acggagcatg
     2641 agaagacgtt cccctgctgc cacctgctct tgtccacggt gagcttgctg tagaggaaga
     2701 aggagccgtc ggagtccagc acgggaggcg tggtcttgta gttgttctcc ggctgcccat
     2761 tgctctccca ctccacggcg atgtcgctgg gatagaagcc tttgaccagg caggtcaggc
     2821 tgacctggtt cttggtcagc tcatcccggg atgggggcag ggtgtacacc tgtggttctc
     2881 ggggctgccc tttggctttg gagatggttt tctcgatggg ggctgggagg gctttgttgg
     2941 agaccttgca cttgtactcc ttgccattca gccagccctg gtgcaggacg gtgaggacgc
     3001 tgaccacacg gtacgtgctg ttgtactgcc cctcccgcgg ctttgtcttg gcattatgca
     3061 cctccacgcc gtccacgtac cagttgaact tgacctcagg gtcttcgtgg ctcacgtcca
     3121 ccaccacgca tgtgacctca ggggtccggg agatcatgag ggtgtccttg ggttttgggg
     3181 ggaagaggaa gactgacggt ccccccagga gttcaggtgc tgggcacggt gggcatgtgt
     3241 gagttttgtc acaagatttg cctccgcctc cgccggatcc agaattcgtg ataactagta
     3301 ggcgttctcc agctcggcct ggttggcttt ggcgctccgg gcgcggggcc gcggcttccg
     3361 gcccttctgc ggccgagcgg cctcggtccc gaagtccttg agctccgact ggttgtggaa
     3421 gcgggtgagg cgcttgcact tgcacgaggc caccaggcgc accttgcgcg cgcgcggcgc
     3481 ctcaccaccg ggacacagca gctgcacgcg ctgcgcgcgg tagcggtcgg ggatgcagcg
     3541 gaagtcgggc ccactaggtc gccaccactt gccgcggccg atggcgttgg gcagcaggcg
     3601 cgccgggccg cactggccgg agcacaccag ctcggtgacc ggcttggcgc tgcggcacgg
     3661 cccatcggtc acgtagcggg tgaagtgcag ctcgcggcag ctgtactcgg acacgtcttt
     3721 ggtctcaaag gggtggtggg gaggccgccc tccgttctcc gcccggttca tggtcttgtt
     3781 gttctccagc tccggtggag gctcggggta ctctccgagc tcggggatga tttccgtggc
     3841 atcattcttg aacgcctgcc acccctggcc ctccactaca cggaaggctg tgtgtaccag
     3901 caggcagacg agacacaggg ccagtgggag ctgcatggtg gaaaatctga attcgtcgac
     3961 aagcttaagt ttaaacgcta gccagcttgg gtctccctat agtgagtcgt attaatttcg
     4021 ataagccagt aagcagtggg ttctctagtt agccagagag ctctgcttat atagacctcc
     4081 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     4141 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     4201 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     4261 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     4321 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     4381 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     4441 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     4501 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     4561 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     4621 taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca
     4681 aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta
     4741 gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac
     4801 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag gggatcggga
     4861 gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4921 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4981 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5041 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5101 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5161 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5221 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5281 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5341 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5401 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5461 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5521 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5581 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5641 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5701 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5761 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5821 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5881 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5941 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6001 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6061 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6121 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6181 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6241 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6301 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6361 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6421 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6481 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6541 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6601 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6661 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6721 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6781 agcgcagcga gtcagtgagc gaggaagcgg aag
//
