LOCUS       sequence-322824-        8085 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 8085)
  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 8085)
  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-1
FEATURES             Location/Qualifiers
     source          1..8085
                     /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             3572..3601
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3617..3634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3678..3695)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3684..3908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3954..4382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4041..4060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4251..4272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4391..4411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4396..4725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4576..4711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4638..4657
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4792..5586
                     /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(4846..4865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5456..5475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5760..5881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5797..5816)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5851..5870
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5930..5946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5930..5946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5943..5965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5954..5970
                     /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(5978..5984,5985..6002,6003..6008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6023..6044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6161..6178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6332..6920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6412..6431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7091..7882,7883..7951))
                     /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     7714..7733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7952..8056)
                     /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 aatgaagtat tcttgctgtg ctctggtttt
      961 ggctgtcctg ggcacagaat tgctgggaag cctctgttcg actgtcagat ccccgaggtt
     1021 cagaggacgg atacagcagg aacgaaaaaa catccgaccc aacattattc ttgtgcttac
     1081 cgatgatcaa gatgtggagc tggggtccct gcaagtcatg aacaaaacga gaaagattat
     1141 ggaacatggg ggggccacct tcatcaatgc ctttgtgact acacccatgt gctgcccgtc
     1201 acggtcctcc atgctcaccg ggaagtatgt gcacaatcac aatgtctaca ccaacaacga
     1261 gaactgctct tccccctcgt ggcaggccat gcatgagcct cggacttttg ctgtatatct
     1321 taacaacact ggctacagaa cagccttttt tggaaaatac ctcaatgaat ataatggcag
     1381 ctacatcccc cctgggtggc gagaatggct tggattaatc aagaattctc gcttctataa
     1441 ttacactgtt tgtcgcaatg gcatcaaaga aaagcatgga tttgattatg caaaggacta
     1501 cttcacagac ttaatcacta acgagagcat taattacttc aaaatgtcta agagaatgta
     1561 tccccatagg cccgttatga tggtgatcag ccacgctgcg ccccacggcc ccgaggactc
     1621 agccccacag ttttctaaac tgtaccccaa tgcttcccaa cacataactc ctagttataa
     1681 ctatgcacca aatatggata aacactggat tatgcagtac acaggaccaa tgctgcccat
     1741 ccacatggaa tttacaaaca ttctacagcg caaaaggctc cagactttga tgtcagtgga
     1801 tgattctgtg gagaggctgt ataacatgct cgtggagacg ggggagctgg agaatactta
     1861 catcatttac accgccgacc atggttacca tattgggcag tttggactgg tcaaggggaa
     1921 atccatgcca tatgactttg atattcgtgt gccttttttt attcgtggtc caagtgtaga
     1981 accaggatca atagtcccac agatcgttct caacattgac ttggccccca cgatcctgga
     2041 tattgctggg ctcgacacac ctcctgatgt ggacggcaag tctgtcctca aacttctgga
     2101 cccagaaaag ccaggtaaca ggtttcgaac aaacaagaag gccaaaattt ggcgtgatac
     2161 attcctagtg gaaagaggca aatttctacg taagaaggaa gaatccagca agaatatcca
     2221 acagtcaaat cacttgccca aatatgaacg ggtcaaagaa ctatgccagc aggccaggta
     2281 ccagacagcc tgtgaacaac cggggcagaa gtggcaatgc attgaggata catctggcaa
     2341 gcttcgaatt cacaagtgta aaggacccag tgacctgctc acagtccggc agagcacgcg
     2401 gaacctctac gctcgcggct tccatgacaa agacaaagag tgcagttgta gggagtctgg
     2461 ttaccgtgcc agcagaagcc aaagaaagag tcaacggcaa ttcttgagaa accaggggac
     2521 tccaaagtac aagcccagat ttgtccatac tcggcagaca cgttccttgt ccgtcgaatt
     2581 tgaaggtgaa atatatgaca taaatctgga agaagaagaa gaattgcaag tgttgcaacc
     2641 aagaaacatt gctaagcgtc atgatgaagg ccacaagggg ccaagagatc tccaggcttc
     2701 cagtggtggc aacaggggca ggatgctggc agatagcagc aacgccgtgg gcccacctac
     2761 cactgtccga gtgacacaca agtgttttat tcttcccaat gactctatcc attgtgagag
     2821 agaactgtac caatcggcca gagcgtggaa ggaccataag gcatacattg acaaagagat
     2881 tgaagctctg caagataaaa ttaagaattt aagagaagtg agaggacatc tgaagagaag
     2941 gaagcctgag gaatgtagct gcagtaaaca aagctattac aataaagaga aaggtgtaaa
     3001 aaagcaagag aaattaaaga gccatcttca cccattcaag gaggctgctc aggaagtaga
     3061 tagcaaactg caacttttca aggagaacaa ccgtaggagg aagaaggaga ggaaggagaa
     3121 gagacggcag aggaaggggg aagagtgcag cctgcctggc ctcacttgct tcacgcatga
     3181 caacaaccac tggcagacag ccccgttctg gaacctggga tctttctgtg cttgcacgag
     3241 ttctaacaat aacacctact ggtgtttgcg tacagttaat gagacgcata attttctttt
     3301 ctgtgagttt gctactggct ttttggagta ttttgatatg aatacagatc cttatcagct
     3361 cacaaataca gtgcacacgg tagaacgagg cattttgaat cagctacacg tacaactaat
     3421 ggagctcaga agctgtcaag gatataagca gtgcaaccca agacctaaga atcttgatgt
     3481 tggaaataaa gatggaggaa gctatgacct acacagagga cagttatggg atggatggga
     3541 aggggatccg agctcggtac caagctttct agaacaaaaa ctcatctcag aagaggatct
     3601 gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttaag
     3661 tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3721 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3781 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3841 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3901 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3961 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4021 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4081 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4141 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4261 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4321 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4381 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4501 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4561 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4621 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4681 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4741 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4801 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4861 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4921 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4981 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5041 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5101 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5161 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5221 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5281 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5341 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5401 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5461 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5521 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5581 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5641 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5701 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5761 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5821 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5881 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5941 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6001 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6061 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6121 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6181 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6241 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7081 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7141 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7201 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7261 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7321 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7381 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7441 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7501 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7561 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7621 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7681 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7741 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7801 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7861 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7921 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7981 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8041 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacg
//
