LOCUS       Exported                7872 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-beta-3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  AUTHORS   Takagi J, Petre BM, Walz T, Springer TA
  TITLE     Global conformational rearrangements in integrin extracellular 
            domains in outside-in and inside-out signaling.
  JOURNAL   Cell. 2002 Sep 6. 110(5):599-11.
  PUBMED    12230977
REFERENCE   2  (bases 1 to 7872)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7872
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             3378..3407
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3423..3440
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3463..3480)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3469..3693
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3739..4167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4036..4057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4176..4196)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4181..4510
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4361..4496
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4423..4442
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4577..5371
                     /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(4631..4650)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5241..5260
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5547..5668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5584..5603)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5638..5657
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5717..5733)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5717..5733)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5730..5752)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5741..5757
                     /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(5765..5795)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5810..5831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5948..5965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6119..6707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6199..6218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6878..7738)
                     /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     7501..7520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7739..7843)
                     /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 gttaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattccg
      961 gcggagcgcc gcgggaggcg gacgagatgc gagcgcggcc gcggccccgg ccgctctggg
     1021 cgactgtgct ggcgctgggg gcgctggcgg gcgttggcgt aggagggccc aacatctgta
     1081 ccacgcgagg tgtgagctcc tgccagcagt gcctggctgt gagccccatg tgtgcctggt
     1141 gctctgatga ggccctgcct ctgggctcac ctcgctgtga cctgaaggag aatctgctga
     1201 aggataactg tgccccagaa tccatcgagt tcccagtgag tgaggcccga gtactagagg
     1261 acaggcccct cagcgacaag ggctctggag acagctccca ggtcactcaa gtcagtcccc
     1321 agaggattgc actccggctc cggccagatg attcgaagaa tttctccatc caagtgcggc
     1381 aggtggagga ttaccctgtg gacatctact acttgatgga cctgtcttac tccatgaagg
     1441 atgatctgtg gagcatccag aacctgggta ccaagctggc cacccagatg cgaaagctca
     1501 ccagtaacct gcggattggc ttcggggcat ttgtggacaa gcctgtgtca ccatacatgt
     1561 atatctcccc accagaggcc ctcgaaaacc cctgctatga tatgaagacc acctgcttgc
     1621 ccatgtttgg ctacaaacac gtgctgacgc taactgacca ggtgacccgc ttcaatgagg
     1681 aagtgaagaa gcagagtgtg tcacggaacc gagatgcccc agagggtggc tttgatgcca
     1741 tcatgcaggc tacagtctgt gatgaaaaga ttggctggag gaatgatgca tcccacttgc
     1801 tggtgtttac cactgatgcc aagactcata tagcattgga cggaaggctg gcaggcattg
     1861 tccagcctaa tgacgggcag tgtcatgttg gtagtgacaa tcattactct gcctccacta
     1921 ccatggatta tccctctttg gggctgatga ctgagaagct atcccagaaa aacatcaatt
     1981 tgatctttgc agtgactgaa aatgtagtca atctctatca gaactatagt gagctcatcc
     2041 cagggaccac agttggggtt ctgtccatgg attccagcaa tgtcctccag ctcattgttg
     2101 atgcttatgg gaaaatccgt tctaaagtag agctggaagt gcgtgacctc cctgaagagt
     2161 tgtctctatc cttcaatgcc acctgcctca acaatgaggt catccctggc ctcaagtctt
     2221 gtatgggact caagattgga gacacggtga gcttcagcat tgaggccaag gtgcgaggct
     2281 gtccccagga gaaggagaag tcctttacca taaagcccgt gggcttcaag gacagcctga
     2341 tcgtccaggt cacctttgat tgtgactgtg cctgccaggc ccaagctgaa cctaatagcc
     2401 atcgctgcaa caatggcaat gggacctttg agtgtggggt atgccgttgt gggcctggct
     2461 ggctgggatc ccagtgtgag tgctcagagg aggactatcg cccttcccag caggacgaat
     2521 gcagcccccg ggagggtcag cccgtctgca gccagcgggg cgagtgcctc tgtggtcaat
     2581 gtgtctgcca cagcagtgac tttggcaaga tcacgggcaa gtactgcgag tgtgacgact
     2641 tctcctgtgt ccgctacaag ggggagatgt gctcaggcca tggccagtgc agctgtgggg
     2701 actgcctgtg tgactccgac tggaccggct actactgcaa ctgtaccacg cgtactgaca
     2761 cctgcatgtc cagcaatggg ctgctgtgca gcggccgcgg caagtgtgaa tgtggcagct
     2821 gtgtctgtat ccagccgggc tcctatgggg acacctgtga gaagtgcccc acctgcccag
     2881 atgcctgcac ctttaagaaa gaatgtgtgg agtgtaagaa gtttgaccgg ggagccctac
     2941 atgacgaaaa tacctgcaac cgttactgcc gtgacgagat tgagtcagtg aaagagctta
     3001 aggacactgg caaggatgca gtgaattgta cctataagaa tgaggatgac tgtgtcgtca
     3061 gattccagta ctatgaagat tctagtggaa agtccatcct gtatgtggta gaagagccag
     3121 agtgtcccaa gggccctgac atcctggtgg tcctgctctc agtgatgggg gccattctgc
     3181 tcattggcct tgccgccctg ctcatctgga aactcctcat caccatccac gaccgaaaag
     3241 aatttgctaa atttgaggaa gaacgcgcca gagcaaaatg ggacacagcc aacaacccac
     3301 tgtataaaga ggccacgtct accttcacca atatcacgta ccggggcact ctcgagcatg
     3361 catctagagg gcccttcgaa caaaaactca tctcagaaga ggatctgaat atgcataccg
     3421 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     3481 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3541 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3601 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3661 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3721 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3781 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3841 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3901 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3961 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4021 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4081 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4141 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4261 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4321 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4381 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4441 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4501 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4561 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4621 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4681 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4741 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4801 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4861 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4921 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4981 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5041 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5101 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5161 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5221 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5281 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5341 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     5401 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5461 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5521 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5581 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5641 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5701 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5761 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5821 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5881 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6541 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6781 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6841 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6901 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6961 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7021 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7081 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7141 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7201 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7261 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7321 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7381 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7441 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7501 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7561 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7621 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7681 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7741 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7801 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7861 tgccacctga cg
//
