LOCUS       Exported                8131 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    hE-cadherin-pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8131)
  AUTHORS   Gottardi CJ, Wong E, Gumbiner BM
  TITLE     E-cadherin suppresses cellular transformation by inhibiting 
            beta-catenin signaling in an adhesion-independent manner.
  JOURNAL   J Cell Biol. 2001 May 28;153(5):1049-60.
  PUBMED    11381089
REFERENCE   2  (bases 1 to 8131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8131
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(993..1853)
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2033..2052)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2224..2603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2604..2807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2758..2778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2852..2871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2852..2870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5673..5691)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5674..5691)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5711..5728)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5717..5941
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5987..6415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6074..6093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6284..6305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6424..6444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6429..6758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6609..6744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6671..6690
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6825..7619
                     /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(6879..6898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7489..7508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7793..7914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7830..7849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7884..7903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7963..7979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7963..7979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7976..7998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7987..8003
                     /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(8011..8041)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8056..8077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     2881 cttgatatcg aattccggaa agcacctgtg agcttgcgga agtcagttca gactccagcc
     2941 cgctccagcc cggcccgacc cgaccgcacc cggcgcctgc cctcgctcgg cgtccccggc
     3001 cagccatggg cccttggagc cgcagcctct cggcgctgct gctgctgctg caggtctcct
     3061 cttggctctg ccaggagccg gagccctgcc accctggctt tgacgccgag agctacacgt
     3121 tcacggtgcc ccggcgccac ctggagagag gccgcgtcct gggcagagtg aattttgaag
     3181 attgcaccgg tcgacaaagg acagcctatt tttccctcga cacccgattc aaagtgggca
     3241 cagatggtgt gattacagtc aaaaggcctc tacggtttca taacccacag atccatttct
     3301 tggtctacgc ctgggactcc acctacagaa agttttccac caaagtcacg ctgaatacag
     3361 tggggcacca ccaccgcccc ccgccccatc aggcctccgt ttctggaatc caagcagaat
     3421 tgctcacatt tcccaactcc tctcctggcc tcagaagaca gaagagagac tgggttattc
     3481 ctcccatcag ctgcccagaa aatgaaaaag gcccatttcc taaaaacctg gttcagatca
     3541 aatccaacaa agacaaagaa ggcaaggttt tctacagcat cactggccaa ggagctgaca
     3601 caccccctgt tggtgtcttt attattgaaa gagaaacagg atggctgaag gtgacagagc
     3661 ctctggatag agaacgcatt gccacataca ctctcttctc tcacgctgtg tcatccaacg
     3721 ggaatgcagt tgaggatcca atggagattt tgatcacggt aaccgatcag aatgacaaca
     3781 agcccgaatt cacccaggag gtctttaagg ggtctgtcat ggaaggtgct cttccaggaa
     3841 cctctgtgat ggaggtcaca gccacagacg cggacgatga tgtgaacacc tacaatgccg
     3901 ccatcgctta caccatcctc agccaagatc ctgagctccc tgacaaaaat atgttcacca
     3961 ttaacaggaa cacaggagtc atcagtgtgg tcaccactgg gctggaccga gagagtttcc
     4021 ctacgtatac cctggtggtt caagctgctg accttcaagg tgaggggtta agcacaacag
     4081 caacagctgt gatcacagtc actgacacca acgataatcc tccgatcttc aatcccacca
     4141 cgtacaaggg tcaggtgcct gagaacgagg ctaacgtcgt aatcaccaca ctgaaagtga
     4201 ctgatgctga tgcccccaat accccagcgt gggaggctgt atacaccata ttgaatgatg
     4261 atggtggaca atttgtcgtc accacaaatc cagtgaacaa cgatggcatt ttgaaaacag
     4321 caaagggctt ggattttgag gccaagcagc agtacattct acacgtagca gtgacgaatg
     4381 tggtaccttt tgaggtctct ctcaccacct ccacagccac cgtcaccgtg gatgtgctgg
     4441 atgtgaatga agcccccatc tttgtgcctc ctgaaaagag agtggaagtg tccgaggact
     4501 ttggcgtggg ccaggaaatc acatcctaca ctgcccagga gccagacaca tttatggaac
     4561 agaaaataac atatcggatt tggagagaca ctgccaactg gctggagatt aatccggaca
     4621 ctggtgccat ttccactcgg gctgagctgg acagggagga ttttgagcac gtgaagaaca
     4681 gcacgtacac agccctaatc atagctacag acaatggttc tccagttgct actggaacag
     4741 ggacacttct gctgatcctg tctgatgtga atgacaacgc ccccatacca gaacctcgaa
     4801 ctatattctt ctgtgagagg aatccaaagc ctcaggtcat aaacatcatt gatgcagacc
     4861 ttcctcccaa tacatctccc ttcacagcag aactaacaca cggggcgagt gccaactgga
     4921 ccattcagta caacgaccca acccaagaat ctatcatttt gaagccaaag atggccttag
     4981 aggtgggtga ctacaaaatc aatctcaagc tcatggataa ccagaataaa gaccaagtga
     5041 ccaccttaga ggtcagcgtg tgtgactgtg aaggggccgc cggcgtctgt aggaaggcac
     5101 agcctgtcga agcaggattg caaattcctg ccattctggg gattcttgga ggaattcttg
     5161 ctttgctaat tctgattctg ctgctcttgc tgtttcttcg gaggagagcg gtggtcaaag
     5221 agcccttact gcccccagag gatgacaccc gggacaacgt ttattactat gatgaagaag
     5281 gaggcggaga agaggaccag gactttgact tgagccagct gcacaggggc ctggacgctc
     5341 ggcctgaagt gactcgtaac gacgttgcac caaccctcat gagtgtcccc cggtatcttc
     5401 cccgccctgc caatcccgat gaaattggaa attttattga tgaaaatctg aaagcggctg
     5461 atactgaccc cacagccccg ccttatgatt ctctgctcgt gtttgactat gaaggaagcg
     5521 gttccgaagc tgctagtctg agctccctga actcctcaga gtcagacaaa gaccaggact
     5581 atgactactt gaacgaatgg ggcaatcgct tcaagaagct ggctgacatg tacggaggcg
     5641 gcgaggacca ctaggggtct agagggccct attctatagt gtcacctaaa tgctagagct
     5701 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5761 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5821 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5881 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     5941 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     6001 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6061 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6181 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6301 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6361 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     6421 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     6481 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6601 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6661 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     6721 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     6781 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6841 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6901 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6961 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     7021 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     7081 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     7141 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     7201 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     7261 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7321 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     7381 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7441 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7501 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7561 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7621 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     7801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     7861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     7921 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     7981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     8041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     8101 tttccagtcg ggaaacctgt cgtgccagct g
//
