LOCUS       sequence-8681-ex        6241 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 6241)
  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 6241)
  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-c
FEATURES             Location/Qualifiers
     source          1..6241
                     /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(1602..1626)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1646..1669
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     1908..1928
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1993..2010)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1999..2223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2269..2697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2356..2375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2566..2587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2706..2726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2711..3040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2891..3026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2953..2972
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3107..3901
                     /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(3161..3180)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3771..3790
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3965..4013
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4054..4073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4173..4195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4233..4251)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4319..4423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4424..4492,4493..5284)
                     /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(4642..4661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5455..6043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5944..5963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6197..6214
                     /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 agcagaatgc
      841 tgtgtcctct gaagaaacca atgactttaa acaagagacc cttccaagta agtccaacga
      901 aagccatgac cacatggatg atatggatga tgaagatgat gatgaccatg tggacagcca
      961 ggactccatt gactcgaacg actctgatga tgtagatgac actgatgatt ctcaccagtc
     1021 tgatgagtct caccattctg atgaatctga tgaactggtc actgattttc ccacggacct
     1081 gccagcaacc gaagttttca ctccagttgt ccccacagta gacacatatg atggccgagg
     1141 tgatagtgtg gtttatggac tgaggtcaaa atctaagaag tttcgcagac ctgacatcca
     1201 gtaccctgat gctacagacg agcacatcac ctcacacatg gaaagcgagg agttgaatgg
     1261 tgcatacaag gccatccccg ttgcccagga cctgaacgcg ccttctgatt gggacagccg
     1321 tgggaaggac agttatgaaa cgagtcagct ggatgaccag agtgctgaag cccacagcca
     1381 caagcagtcc agattatata agcggaaagc taatgatgag agcaatgagc attccgatgt
     1441 gattgatagt caggaacttt ccaaagtcag ccgtgaattc cacagccatg aatttcacag
     1501 ccatgaagat atgctggttg tagaccccaa aagtaaggaa gaagataaac acctgaaatt
     1561 tcgtatttct catgaattag atagtgcatc ttctgaggtc aatccaactt tcttgtacaa
     1621 agtggttcga tctagaatgg ctagcgacta caaggacgac gatgacaagt agtaatcgag
     1681 tacaactata actcacacaa tgtatacatc acggcagaca aacaaaagaa tggaatcaaa
     1741 gctaacttca aaattcgcca caacattgaa gatggatccg ttcaactagc agaccattat
     1801 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcg
     1861 acacaatctg ccctttcgaa agatcccaac gaaaagcgtg accacatggt ccttcttgag
     1921 tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaata atgaattaaa
     1981 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2041 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2101 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2161 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2221 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2281 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2341 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2401 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2461 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2521 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2581 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2641 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2701 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2761 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2821 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2881 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2941 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3001 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3061 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3121 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3181 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3241 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3301 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3361 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3421 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3481 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3541 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3601 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3661 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3721 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3781 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3841 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3901 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     3961 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4021 agcgataagg atccgcgtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4081 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4141 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4201 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4261 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4321 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4381 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4441 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4501 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4561 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4621 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4681 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4741 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4801 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4861 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4921 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4981 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5041 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5101 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5161 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5221 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5281 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5341 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5401 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5461 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5521 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5581 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5641 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5701 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5761 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5821 cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa
     5881 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5941 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6001 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6061 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6121 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6181 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagctcg cgaatgcatg
     6241 t
//
