LOCUS       Exported                9607 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pHAHA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9607)
  AUTHORS   Luu HH, Zhou L, Haydon RC, Deyrup AT, Montag AG, Huo D, Heck R, 
            Heizmann CW, Peabody TD, Simon MA, He TC
  TITLE     Increased expression of S100A6 is associated with decreased 
            metastasis and inhibition of cell migration and anchorage 
            independent growth in human osteosarcoma.
  JOURNAL   Cancer Lett. 2005 Nov 8. 229(1):135-48.
  PUBMED    16157226
REFERENCE   2  (bases 1 to 9607)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9607
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        141..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     721..745
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    852..986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(876..895)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     930..949
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1129..1151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      1711..3500
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(5538..5556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5624..5728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5729..6589
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5947..5966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6760..7348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7249..7268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7779..7924
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             7967..9004
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(9002..9021)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    9046..9094
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
       61 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
      121 tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
      661 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 agctcgttta gtgaaccgtc agatctctag aagctgggta ccgaaatggg atacgatgtt
      781 ccagattacg cttcgtacga tgttccagat tacgcaagct tgctagcggc cgctcgaggc
      841 cggcaaggcc ggatccagac atgataagat acattgatga gtttggacaa accacaacta
      901 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      961 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1021 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     1081 ctgattatga tccggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1141 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1201 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaggt cgactctaga
     1261 ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc
     1321 ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc
     1381 acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt
     1441 ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca
     1501 gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac
     1561 caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac
     1621 gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat
     1681 agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag
     1741 tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg
     1801 ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag
     1861 ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc
     1921 acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc
     1981 ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt
     2041 atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg
     2101 ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat
     2161 aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg
     2221 gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca
     2281 taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac
     2341 aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag
     2401 cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat
     2461 ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac
     2521 gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt
     2581 ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg
     2641 gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg
     2701 cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc
     2761 ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac
     2821 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc
     2881 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     2941 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     3001 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     3061 ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc
     3121 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa
     3181 tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     3241 tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa
     3301 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     3361 tctgtatccg ggtagcatat gctatcctca tgcatataca gtcagcatat gatacccagt
     3421 agtagagtgg gagtgctatc ctttgcatat gccgccacct cccaaggggg cgtgaatttt
     3481 cgctgcttgt ccttttcctg ctggttgctc ccattcttag gtgaatttaa ggaggccagg
     3541 ctaaagccgt cgcatgtctg attgctcacc aggtaaatgt cgctaatgtt ttccaacgcg
     3601 agaaggtgtt gagcgcggag ctgagtgacg tgacaacatg ggtatgccca attgccccat
     3661 gttgggagga cgaaaatggt gacaagacag atggccagaa atacaccaac agcacgcatg
     3721 atgtctactg gggatttatt ctttagtgcg ggggaataca cggcttttaa tacgattgag
     3781 ggcgtctcct aacaagttac atcactcctg cccttcctca ccctcatctc catcacctcc
     3841 ttcatctccg tcatctccgt catcaccctc cgcggcagcc ccttccacca taggtggaaa
     3901 ccagggaggc aaatctactc catcgtcaaa gctgcacaca gtcaccctga tattgcaggt
     3961 aggagcgggc tttgtcataa caaggtcctt aatcgcatcc ttcaaaacct cagcaaatat
     4021 atgagtttgt aaaaagacca tgaaataaca gacaatggac tcccttagcg ggccaggttg
     4081 tgggccgggt ccaggggcca ttccaaaggg gagacgactc aatggtgtaa gacgacattg
     4141 tggaatagca agggcagttc ctcgccttag gttgtaaagg gaggtcttac tacctccata
     4201 tacgaacaca ccggcgaccc aagttccttc gtcggtagtc ctttctacgt gactcctagc
     4261 caggagagct cttaaacctt ctgcaatgtt ctcaaatttc gggttggaac ctccttgacc
     4321 acgatgcttt ccaaaccacc ctcctttttt gcgcctgcct ccatcaccct gaccccgggg
     4381 tccagtgctt gggccttctc ctgggtcatc tgcggggccc tgctctatcg ctcccggggg
     4441 cacgtcaggc tcaccatctg ggccaccttc ttggtggtat tcaaaataat cggcttcccc
     4501 tacagggtgg aaaaatggcc ttctacctgg agggggcctg cgcggtggag acccggatga
     4561 tgatgactga ctactgggac tcctgggcct cttttctcca cgtccacgac ctctccccct
     4621 ggctctttca cgacttcccc ccctggctct ttcacgtcct ctaccccggc ggcctccact
     4681 acctcctcga ccccggcctc cactacctcc tcgaccccgg cctccactgc ctcctcgacc
     4741 ccggcctcca cctcctgctc ctgcccctcc tgctcctgcc cctcctcctg ctcctgcccc
     4801 tcctgctcct gcccctcccg ctcctgctcc tgctcctgtt ccaccgtggg tccctttgca
     4861 gccaatgcaa cttggacgtt tttggggtct ccggacacca tctctatgtc ttggccctga
     4921 tcctgagccg cccggggctc ctggtcttcc gcctcctcgt cctcgtcctc ttccccgtcc
     4981 tcgtccatgg ttatcacccc ctcttctttg aggtccactg ccgccggagc cttctggtcc
     5041 agatgtgtct cccttctctc ctaggccatt tccaggtcct gtacctggcc cctcgtcaga
     5101 catgattcac actaaaagag atcaatagac atctttatta gacgacgctc agtgaataca
     5161 gggagtgcag actcctgccc cctccaacag cccccccacc ctcatcccct tcatggtcgc
     5221 tgtcagacag atccaggtct gaaaattccc catcctccga accatcctcg tcctcatcac
     5281 caattactcg cagcccggaa aactcccgct gaacatcctc aagatttgcg tcctgagcct
     5341 caagccaggc ctcaaattcc tcgtccccct ttttgctgga cggtagggat ggggattctc
     5401 gggacccctc ctcttcctct tcaaggtcac cagacagaga tgctactggg gcaacggaag
     5461 aaaagctggg tgcggcctgt gaggatcagc ttatcgatga taagctgtca aacatgagaa
     5521 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5581 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5641 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5701 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5761 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5821 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5881 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5941 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     6001 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6061 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6121 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6181 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6241 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     6301 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6361 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6421 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6481 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6541 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6601 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6661 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6721 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6781 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6841 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6901 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6961 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7021 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7081 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7141 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7201 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7261 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7321 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7381 ggccttttgc tggccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
     7441 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
     7501 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccccgagatg
     7561 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     7621 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     7681 ttagcgaggt gccgccctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt
     7741 ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg
     7801 tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt ccgaggtcca
     7861 cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc
     7921 ttaacagcgt caacagcgtg ccgcagatcc cggggggcaa tgagatatga aaaagcctga
     7981 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     8041 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     8101 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     8161 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     8221 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     8281 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     8341 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     8401 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     8461 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     8521 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     8581 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     8641 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     8701 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     8761 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     8821 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     8881 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     8941 cgccgatagt ggaaaccgac gccccagcac tcgtccggat cgggagatgg gggaggctaa
     9001 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     9061 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     9121 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     9181 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     9241 gcggcaggcc ctgccatagc cactggcccc gtgggttagg gacggggtcc cccatgggga
     9301 atggtttatg gttcgtgggg gttattattt tgggcgttgc gtggggtcag gtccacgact
     9361 ggactgagca gacagaccca tggtttttgg atggcctggg catggaccgc atgtactggc
     9421 gcgacacgaa caccgggcgt ctgtggctgc caaacacccc cgacccccaa aaaccaccgc
     9481 gcggatttct ggcgtgccaa gctagtcgac caattctcat gtttgacagc ttatcatcgc
     9541 agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct
     9601 atacatt
//
