LOCUS       sequence-208115-        8064 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 8064)
  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 8064)
  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(-)-HSulf-2
FEATURES             Location/Qualifiers
     source          1..8064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3549..3578
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3594..3611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3655..3672)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3661..3885
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3931..4359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4018..4037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4228..4249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4368..4388)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4373..4702
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4553..4688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4615..4634
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4769..5563
                     /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(4823..4842)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5433..5452
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5737..5858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5774..5793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5828..5847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5907..5923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5907..5923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5920..5942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5931..5947
                     /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(5955..5961,5962..5979,5980..5985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6000..6021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6138..6155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6309..6897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6389..6408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7068..7859,7860..7928))
                     /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     7691..7710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7929..8033)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagaaga tgggcccccc gagcctcgtg ctgtgcttgc
      961 tgtccgcaac tgtgttctcc ctgctgggtg gaagctcggc cttcctgtcg caccaccgcc
     1021 tgaaaggcag gtttcagagg gaccgcagga acatccgccc caacatcatc ctggtgctga
     1081 cggacgacca ggatgtggag ctgggttcca tgcaggtgat gaacaagacc cggcgcatca
     1141 tggagcaggg cggggcgcac ttcatcaacg ccttcgtgac cacacccatg tgctgcccct
     1201 cacgctcctc catcctcacc ggcaagtacg tccacaacca caacacctac accaacaatg
     1261 agaactgctc ctcgccctcc tggcaggcac agcacgagag ccgcaccttt gccgtgtacc
     1321 tcaatagcac tggctaccgg acagctttct tcgggaagta tcttaatgaa tacaacggct
     1381 cctacgtgcc acccggctgg aaggagtggg tcggactcct taaaaactcc cgcttttata
     1441 actacacgct gtgtcggaac ggggtgaaag agaagcacgg ctccgactac tccaaggatt
     1501 acctcacaga cctcatcacc aatgacagcg tgagcttctt ccgcacgtcc aagaagatgt
     1561 acccgcacag gccagtcctc atggtcatca gccatgcagc cccccacggc cctgaggatt
     1621 cagccccaca atattcacgc ctcttcccaa acgcatctca gcacatcacg ccgagctaca
     1681 actacgcgcc caacccggac aaacactgga tcatgcgcta cacggggccc atgaagccca
     1741 tccacatgga attcaccaac atgctccagc ggaagcgctt gcagaccctc atgtcggtgg
     1801 acgactccat ggagacgatt tacaacatgc tggttgagac gggcgagctg gacaacacgt
     1861 acatcgtata caccgccgac cacggttacc acatcggcca gtttggcctg gtgaaaggga
     1921 aatccatgcc atatgagttt gacatcaggg tcccgttcta cgtgaggggc cccaacgtgg
     1981 aagccggctg tctgaatccc cacatcgtcc tcaacattga cctggccccc accatcctgg
     2041 acattgcagg cctggacata cctgcggata tggacgggaa atccatcctc aagctgctgg
     2101 acacggagcg gccggtgaat cggtttcact tgaaaaagaa gatgagggtc tggcgggact
     2161 ccttcttggt ggagagaggc aagctgctac acaagagaga caatgacaag gtggacgccc
     2221 aggaggagaa ctttctgccc aagtaccagc gtgtgaagga cctgtgtcag cgtgctgagt
     2281 accagacggc gtgtgagcag ctgggacaga agtggcagtg tgtggaggac gccacgggga
     2341 agctgaagct gcataagtgc aagggcccca tgcggctggg cggcagcaga gccctctcca
     2401 acctcgtgcc caagtactac gggcagggca gcgaggcctg cacctgtgac agcggggact
     2461 acaagctcag cctggccgga cgccggaaaa aactcttcaa gaagaagtac aaggccagct
     2521 atgtccgcag tcgctccatc cgctcagtgg ccatcgaggt ggacggcagg gtgtaccacg
     2581 taggcctggg tgatgccgcc cagccccgaa acctcaccaa gcggcactgg ccaggggccc
     2641 ctgaggacca agatgacaag gatggtgggg acttcagtgg cactggaggc cttcccgact
     2701 actcagccgc caaccccatt aaagtgacac atcggtgcta catcctagag aacgacacag
     2761 tccagtgtga cctggacctg tacaagtccc tgcaggcctg gaaagaccac aagctgcaca
     2821 tcgaccacga gattgaaacc ctgcagaaca aaattaagaa cctgagggaa gtccgaggtc
     2881 acctgaagaa aaagcggcca gaagaatgtg actgtcacaa aatcagctac cacacccagc
     2941 acaaaggccg cctcaagcac agaggctcca gtctgcatcc tttcaggaag ggcctgcaag
     3001 agaaggacaa ggtgtggctg ttgcgggagc agaagcgcaa gaagaaactc cgcaagctgc
     3061 tcaagcgcct gcagaacaac gacacgtgca gcatgccagg cctcacgtgc ttcacccacg
     3121 acaaccagca ctggcagacg gcgcctttct ggacactggg gcctttctgt gcctgcacca
     3181 gcgccaacaa taacacgtac tggtgcatga ggaccatcaa tgagactcac aatttcctct
     3241 tctgtgaatt tgcaactggc ttcctagagt actttgatct caacacagac ccctaccagc
     3301 tgatgaatgc agtgaacaca ctggacaggg atgtcctcaa ccagctacac gtacagctca
     3361 tggagctgag gagctgcaag ggttacaagc agtgtaaccc ccggactcga aacatggacc
     3421 tgggacttaa agatggagga agctatgagc aatacaggca gtttcagcgt cgaaagtggc
     3481 cagaaatgaa gagaccttct tccaaatcac tgggacaact gtgggaaggc tgggaaggaa
     3541 gctttctaga acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc
     3601 atcatcatca ttgagtttaa acggtctcca gcttaagttt aaaccgctga tcagcctcga
     3661 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3721 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3781 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3841 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3901 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3961 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4021 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4081 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4141 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4201 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4261 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4321 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4381 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4441 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4501 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4561 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4621 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4681 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4741 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4801 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4861 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4921 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4981 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5041 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5101 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5161 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5221 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5281 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5341 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5401 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5461 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5521 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5581 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5641 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5701 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5821 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5881 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5941 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6001 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6061 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6121 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6181 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6241 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6481 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6721 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6781 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7081 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7141 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7201 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7261 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7321 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7381 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7441 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7501 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7561 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7621 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7681 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7741 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7801 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7861 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7921 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8041 ccccgaaaag tgccacctga cgtc
//
