LOCUS       sequence-207636-        8079 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 8079)
  AUTHORS   Morimoto-Tomita M, Uchimura K, Werb Z, Hemmerich S, Rosen SD
  TITLE     Cloning and characterization of two extracellular heparin-degrading 
            endosulfatases in mice and humans.
  JOURNAL   J Biol Chem. 2002 Dec 20. 277(51):49175-85.
  PUBMED    12368295
REFERENCE   2  (bases 1 to 8079)
  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: "J Biol 
            Chem. 2002 Dec 20. 277(51):49175-85."
COMMENT     Sequence Label: pcDNA3.1/Myc-His(-)-MSulf-2
FEATURES             Location/Qualifiers
     source          1..8079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3566..3595
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3611..3628
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3672..3689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3678..3902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3948..4376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4245..4266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4385..4405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4390..4719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4570..4705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4632..4651
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4786..5580
                     /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(4840..4859)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5450..5469
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5754..5875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5791..5810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5845..5864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5924..5940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5924..5940)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5937..5959)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5948..5964
                     /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(5972..5978,5979..5996,5997..6002))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6017..6038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6155..6172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6326..6914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6406..6425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7085..7876,7877..7945))
                     /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     7708..7727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7946..8050)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac gggccctcta gactcgagac aatggcaccc cctggcctgc cactatggct
      961 gctgtccacc gctctcctct ccctgctggc tggcagctcg gccttcctct cccatccccg
     1021 cctgaaggga cgcttccaga gggaccgcag gaacatccgg cccaacatca tcttggtgct
     1081 tacggatgac caggatgtgg agctgggctc catgcaagtg atgaacaaga caaggcgtat
     1141 catggagcag ggcggggcgc acttcatcaa tgccttcgtg actacaccaa tgtgctgtcc
     1201 gtctcgctcc tccattctca ccggcaagta cgtccacaac cacaacacct acaccaacaa
     1261 tgagaattgt tcctcgccct cctggcaggc ccagcacgag agccgcacct tcgccgtgta
     1321 tctcaacagc acaggctacc ggacagcttt cttcggaaaa tacctcaatg agtacaacgg
     1381 ctcatacgtg ccgcccggct ggaaggagtg ggtcggccta cttaagaact cccgctttta
     1441 taactacaca ctctgccgga atggggtgaa ggagaaacat ggctcggact actccacgga
     1501 ttacctcacg gatctcatca ccaatgacag tgtgagcttc ttccgaacat ccaagaagat
     1561 gtacccacac aggcccgtgc tcatggtcat cagccacgcg gctccccatg gccccgagga
     1621 ctcagcaccc cagtactcac ggctcttccc caatgcgtcc cagcacatca caccgagtta
     1681 caactatgca cccaacccag acaagcattg gatcatgcgc tacacgggac ccatgaagcc
     1741 cattcacatg gaattcacca acatgctaca acgaaaacgc ctacagaccc tcatgtctgt
     1801 ggatgactcc atggagacga tctatgacat gctggtggag acgggggagc tggacaacac
     1861 gtacatcctg tacaccgccg accacggcta ccacattggc cagtttgggc tggtgaaggg
     1921 caagtctatg ccgtatgaat tcgacatcag agtcccgttc tacgtgaggg gccccaacgt
     1981 ggaagctggc tctctgaacc cccacattgt cctcaacatt gacctggccc ccaccatact
     2041 ggatatcgct ggactggaca tccctgcaga catggacggg aagtctattc tcaaactact
     2101 ggactcagag cggccagtga accggttcca cttgaaaaag aagctgaggg tctggcgaga
     2161 ctccttcctg gtggagagag gcaaactgct ccacaagagg gagggtgaca aagtgaatgc
     2221 ccaggaggag aacttcctgc ccaagtacca gcgcgtgaag gacctgtgtc agcgagctga
     2281 gtaccagaca gcatgcgaac agctggggca gaagtggcag tgtgtggagg acgcttctgg
     2341 gacgctgaag ctgcacaaat gtaaaggccc catgcggttt ggtggcggcg gtggcagcag
     2401 agccctctcc aacctggtgc ccaagtatga cggccagagc agcgaggcct gcagctgtga
     2461 cagtggcggt ggaggggact acaaactggg cctggctgga cgccgtaagc tctttaagaa
     2521 aaagtataag accagctatg cccggaaccg ctccatccgt tccgtggcca tcgaggtgga
     2581 cggtgagata taccacgtag gcttggatac tgtacctcag ccccgcaacc ttagcaagcc
     2641 gcactggcca ggggcccctg aagaccaaga tgacaaggat ggtggcagtt tcagtggtac
     2701 tggtggcctt ccagattatt ctgcccccaa tcccatcaaa gtgacccatc ggtgctacat
     2761 ccttgagaat gacacagtcc agtgcgactt ggacctgtac aagtccctgc aggcttggaa
     2821 agaccacaag ctgcacatcg accatgagat cgaaaccctg cagaacaaaa ttaagaacct
     2881 tcgagaagtc aggggtcacc tgaagaagaa gcgaccggaa gaatgtgact gccataaaat
     2941 cagttaccac agccaacaca aaggccgtct caagcacaaa ggctccagcc tgcacccttt
     3001 caggaagggt ctgcaggaga aggacaaggt gtggctgctg cgggagcaga aacgcaagaa
     3061 gaaactgcgc aagctgctca aacggctgca gaacaacgat acgtgcagca tgcccggcct
     3121 cacgtgcttt acccacgaca accaccactg gcagacggcg ccactctgga cgctggggcc
     3181 gttctgcgcc tgcaccagcg ccaacaacaa cacgtactgg tgcttgagga ccataaatga
     3241 gacccacaac ttcctcttct gcgaatttgc aaccggcttc atagaatact ttgacctcag
     3301 tacagacccc taccagctga tgaacgcggt gaacacactg gacagggacg tccttaacca
     3361 actgcacgtg cagctcatgg agctaaggag ctgtaaaggc tacaagcagt gcaacccccg
     3421 gacccgcaac atggacctgg ggcttagaga cggaggaagc tatgaacaat acaggcagtt
     3481 tcagcgtcga aaatggccag aaatgaagag accttcttcc aaatcactgg gacagctatg
     3541 ggaaggctgg gaaggaagct ttctagaaca aaaactcatc tcagaagagg atctgaatag
     3601 cgccgtcgac catcatcatc atcatcattg agtttaaacg gtctccagct taagtttaaa
     3661 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3721 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3781 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3841 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3901 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3961 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4021 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4081 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4141 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4201 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4261 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4321 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4381 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4441 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4561 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4621 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4681 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4741 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4801 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4861 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4921 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4981 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5041 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5101 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5161 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5221 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5281 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5341 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5401 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5461 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5521 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5581 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5641 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5701 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5761 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5821 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5881 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5941 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6001 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6061 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6121 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6181 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6241 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6301 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6361 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6421 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6481 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6541 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6601 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6661 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6721 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6781 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6841 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6901 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6961 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7021 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7081 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7141 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7201 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7261 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7321 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7381 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7441 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7501 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7561 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7621 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7681 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7741 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7801 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7861 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7921 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7981 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8041 gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
