LOCUS       sequence-238567-        6280 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 6280)
  AUTHORS   Takafuji V, Forgues M, Unsworth E, Goldsmith P, Wang XW
  TITLE     An osteopontin fragment is essential for tumor cell invasion in 
            hepatocellular carcinoma.
  JOURNAL   Oncogene. 2007 Sep 27. 26(44):6361-71.
  PUBMED    17452979
REFERENCE   2  (bases 1 to 6280)
  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: "Oncogene. 
            2007 Sep 27. 26(44):6361-71."
COMMENT     Sequence Label: pDest490-OPN-b
FEATURES             Location/Qualifiers
     source          1..6280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     634..650
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     634..650
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        673..691
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     673..690
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    716..740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1641..1665)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1685..1708
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     1947..1967
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2032..2049)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2038..2262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2308..2736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2395..2414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2605..2626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2745..2765)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2750..3079
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2930..3065
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2992..3011
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3146..3940
                     /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(3200..3219)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3810..3829
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4004..4052
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4093..4112)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4212..4234
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4272..4290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4358..4462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4463..4531,4532..5323)
                     /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(4681..4700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5494..6082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5983..6002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6236..6253
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agcctaggcc gcgggacgga taacaatttc acacaggaaa cagctatgac cattaggcct
      661 ttgcaaaaag ctatttaggt gacactatag aaggtacgcc tgcaggtacc ggatcacaag
      721 tttgtacaaa aaagttggca ccatgagaat tgcagtgatt tgcttttgcc tcctaggcat
      781 cacctgtgcc ataccagtta aacaggctga ttctggaagt tctgaggaaa agcagcttta
      841 caacaaatac ccagatgctg tggccacatg gctaaaccct gacccatctc agaagcagaa
      901 tctcctagcc ccacagaccc ttccaagtaa gtccaacgaa agccatgacc acatggatga
      961 tatggatgat gaagatgatg acgaccatgt ggacagccag gactccattg actcgaacga
     1021 ctctgatgat gtagatgaca ctgatggttc tcaccagtct gatgagtctc accattctga
     1081 tgaatctgat gaactggtca ctgattttcc cacggacctg ccagcaaccg aagttttcac
     1141 tccagttgtc cccacagtag acacatatga tggccgaggt gatagtgtgg tttatggact
     1201 gaggtcaaaa tctaagaagt ttcgcagacc tgacatccag taccctgatg ctacagacga
     1261 ggacatcacc tcacacatgg aaagcgagga gttgaatggt gcatacaagg ccatccccgt
     1321 tgcccaggac ctgaacgcgc cttctgattg ggacagccgt gggaaggaca gttatgaaac
     1381 gagtcagctg gatgaccaga gtgctgaaac ccacagccac aagcagtcca gattatataa
     1441 gcggaaagct aatgatgaga gcaatgagca ttccgatgtg attgatagtc aggaactttc
     1501 caaagtcagc cgtgaattcc acagccatga atttcacagc catgaagata tgctggttgt
     1561 agaccccaaa agtaaggaag aagataaaca cctgaaattt cgtatttctc atgaattaga
     1621 tagtgcatct tctgaggtca atccaacttt cttgtacaaa gtggttcgat ctagaatggc
     1681 tagcgactac aaggacgacg atgacaagta gtaatcgagt acaactataa ctcacacaat
     1741 gtatacatca cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac
     1801 aacattgaag atggatccgt tcaactagca gaccattatc aacaaaatac tccaattggc
     1861 gatggccctg tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa
     1921 gatcccaacg aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt
     1981 acacatggca tggatgagct ctacaaataa tgaattaaac ccgctgatca gcctcgactg
     2041 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2101 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2161 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2221 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2281 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2341 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2401 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2461 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2521 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2581 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2641 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2701 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2761 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2821 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2881 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2941 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3001 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3061 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3121 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3181 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3241 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3301 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3361 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3421 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3481 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3541 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3601 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3661 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3721 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3781 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3841 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3901 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     3961 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     4021 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg
     4081 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4141 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4201 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4261 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4321 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4381 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4441 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4501 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4561 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4621 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4681 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4741 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4801 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4861 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4921 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4981 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5041 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5101 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5161 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5221 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5281 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5641 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6181 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6241 gtcagtgagc gaggaagcgg aagagctcgc gaatgcatgt
//
