LOCUS       sequence-183013-        9069 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 9069)
  AUTHORS   Wei X, Moncada-Pazos A, Cal S, Soria-Valles C, Gartner J, Rudloff U,
            Lin JC, Rosenberg SA, Lopez-Otin C, Samuels Y
  TITLE     Analysis of the disintegrin-metalloproteinases family reveals ADAM29
            and ADAM7 are often mutated in melanoma.
  JOURNAL   Hum Mutat. 2011 Jun;32(6):E2148-75. doi: 10.1002/humu.21477. Epub 
            2011 Feb 24.
  PUBMED    21618342
REFERENCE   2  (bases 1 to 9069)
  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"; doi: 
            "10.1002/humu.21477"; journalName: "Hum Mutat"; date: "2011-06"; 
            volume: "32"; issue: "6"; pages: "E2148-75"
COMMENT     Sequence Label: pCDF1-ADAM29 (wt)
FEATURES             Location/Qualifiers
     source          1..9069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1274..1477
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1474..1498
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4014..4037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        4082..4293
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4306..4574
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4601..5200
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4601..4620)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5097..5117
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5207..5795
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5260..5280)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5678..5689)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6132..6253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6169..6188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6223..6242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6293..6428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6461..6477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6461..6477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6474..6496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6485..6501
                     /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(join(6509..6515,6516..6533,6534..6539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6554..6575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6692..6709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6863..7451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6943..6962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7622..8413,8414..8482))
                     /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     8245..8264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8483..8587)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8655..8673
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8711..8733)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8833..8852
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(8997..9069,1..131)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
       61 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      121 taagcagagc ttgtgaaact tcgaggagtc tctttgttga ggacttttga gttctccctt
      181 gaggctccca cagatacaat aaatatttga gattgaaccc tgtcgagtat ctgtgtaatc
      241 ttttttacct gtgaggtctc ggaatccggg ccgagaactt cgcagttggc gcccgaacag
      301 ggacttgatt gagagtgatt gaggaagtga agctagagca atagaaagct gttaagcaga
      361 actcctgctg acctaaatag ggaagcagta gcagacgctg ctaacagtga gtatctctag
      421 tgaagcagac tcgagctcat aatcaagtca ttgtttaaag gcccagataa attacatctg
      481 gtgactcttc gcggaccttc aagccaggag attcgccgag ggacagtcaa caaggtagga
      541 gagattctac agcaacatgg ggaatggaca ggggcgagat tggaaaatgg ccattaagag
      601 atgtagtaat gttgctgtag gagtaggggg gaagagtaaa aaatttggag aagggaattt
      661 cagatgggcc attagaatgg ctaatgtatc tacaggacga gaacctggtg atataccaga
      721 gactttagat caactaaggt tggttatttg cgatttacaa gaaagaagag aaaaatttgg
      781 atctagcaaa gaaattgata tggcaattcc tgcattgagg agaaatggta ggcaatgtgg
      841 catgtctgaa aaagaggagg aatgatgaag tatctcagac ttattttata agggagatac
      901 tgtgctgagt tcttcccttt gaggaaggta tgtcatatcc tagacatagt ctcaatttta
      961 aaagaagagg taggatagga gggatggccc cttatgaatt attagcacaa caagaatcct
     1021 taagaataca agattatttt tctgcaatac cacaaaaatt gcaagcacag tggatttatt
     1081 ataaagatca aaaagataag aaatggaaag gaccaatgag agtagaatac tggggacagg
     1141 gatcagtatt attaaaggat gaagagaagg gatattttct tataatcgat actagtatta
     1201 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1261 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     1321 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1381 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1441 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     1501 ctggagacgc catccacgct gttttgacct ccatagaaga ttctagagcc accatgaaga
     1561 tgttactcct gctgcattgc cttggggtgt ttctgtcctg ttctggacac atccaggatg
     1621 agcaccccca atatcacagc cctccggatg tggtgattcc tgtgaggata actggcacca
     1681 ccagaggcat gacacctcca ggctggctct cctatatcct gccctttgga ggccagaaac
     1741 acattatcca cataaaggtc aagaagcttt tgttttccaa acacctccct gtgttcacct
     1801 acacagacca gggtgctatc cttgaggacc agccatttgt ccagaataac tgctactatc
     1861 atggttatgt ggaaggggac ccagaatccc tggtttccct cagtacctgt tttgggggtt
     1921 ttcaaggaat attacagata aatgactttg cttatgaaat caagccccta gcattttcta
     1981 ccacgtttga acatctggta tacaagatgg acagtgagga gaaacaattt tcaaccatga
     2041 gatccggatt tatgcaaaat gaaataacat gccgaatgga atttgaagaa attgataatt
     2101 ccactcagaa gcaaagttct tatgtgggct ggtggatcca ttttaggatt gttgaaattg
     2161 tagtcgtcat tgataattat ctgtacattc gttatgaaag gaacgactca aagttgctgg
     2221 aggatctata tgttattgtt aatatagtgg attccatttt ggatgtcatt ggtgttaagg
     2281 tgttattatt tggtttggag atctggacca ataaaaacct cattgtagta gatgatgtaa
     2341 ggaaatctgt gcacctgtat tgcaagtgga agtcggagaa cattacgccc cggatgcaac
     2401 atgacacctc acatcttttc acaactctag gattaagagg gttaagtggc ataggagctt
     2461 ttagaggaat gtgtacacca caccgtagtt gtgcaattgt tactttcatg aacaaaactt
     2521 tgggcacttt ttcaattgca gtggctcatc atctaggtca taatttgggc atgaaccatg
     2581 atgaggatac atgtcgttgt tcacaaccta gatgcataat gcatgaaggc aacccaccaa
     2641 taactaaatt tagcaattgt agttatggtg atttttggga atatactgta gagaggacaa
     2701 agtgtttgct tgaaacagta cacacaaagg acatctttaa tgtgaagcgc tgtgggaatg
     2761 gtgttgttga agaaggagaa gagtgtgact gtggaccttt aaagcattgt gcaaaagatc
     2821 cctgctgtct gtcaaattgc actctgactg atggttctac ttgtgctttt gggctttgtt
     2881 gcaaagactg caagttccta ccatcaggga aagtgtgtag aaaggaggtc aatgaatgtg
     2941 atcttccaga gtggtgcaat ggtacttccc ataagtgccc agatgacttt tatgtggaag
     3001 atggaattcc ctgtaaggag aggggctact gctatgaaaa gagctgtcat gaccgcaatg
     3061 aacagtgtag gaggattttt ggtgcaggcg caaatactgc aagtgagact tgctacaaag
     3121 aattgaacac cttaggtgac cgtgttggtc actgtggtat caaaaatgct acatatataa
     3181 agtgtaatat ctcagatgtc cagtgtggaa gaattcagtg tgagaatgtg acagaaattc
     3241 ccaatatgag tgatcatact actgtgcatt gggctcgctt caatgacata atgtgctgga
     3301 gtactgatta ccatttgggg atgaagggac ctgatattgg tgaagtgaaa gatggaacag
     3361 agtgtgggat agatcatata tgcatccaca ggcactgtgt ccatataacc atcttgaata
     3421 gtaattgctc acctgcattt tgtaacaaga ggggcatctg caacaataaa catcactgcc
     3481 attgcaatta tctgtgggac cctcccaact gcctgataaa aggctatgga ggtagtgttg
     3541 acagtggccc accccctaag agaaagaaga aaaagaagtt ctgttatctg tgtatattgt
     3601 tgcttattgt tttgtttatt ttattatgtt gtctttatcg actttgtaaa aaaagtaaac
     3661 caataaaaaa gcagcaagat gttcaaactc catctgcaaa agaagaggaa aaaattcagc
     3721 gtcgacctca tgagttacct ccccagagtc aaccttgggt gatgccttcc cagagtcaac
     3781 ctcctgtgac gccttcccag agtcatcctc aggtgatgcc ttcccagagt caacctcctg
     3841 tgacaccctc ccagagtcaa cctcgggtga tgccttctca gagtcaacct cctgtgatgc
     3901 cttcccagag tcatcctcag ttgacgcctt cccagagtca acctcctgtg acaccctccc
     3961 agaggcaacc tcagttgatg ccttcccaga gtcaacctcc tgtgacgccc tccgattaca
     4021 aggatgacga cgataagtag gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4081 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4141 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4201 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4261 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4321 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4381 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4441 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4501 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4561 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     4621 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4681 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4741 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4801 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4861 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4921 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4981 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5041 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5101 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5161 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5341 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5401 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5461 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5521 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5581 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     5641 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5701 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5761 gacgagtcgg atctcccttt gggccgcctc cccgcctggt accgatgaca gagttagaag
     5821 atcgcttcag gaagctattt ggcacgactt ctacaacggg agacagcaca gtagattctg
     5881 aagatgaacc tcctaaaaaa gaaaaaaggg tggactggga tgagtattgg aaccctgaaa
     5941 tcgatagctt ccagtgcttt gtgaaacttc gaggagtctc tttgttgagg acttttgagt
     6001 tctcccttga ggctcccaca gatacaataa atatttgaga ttgaaccctg tcgagtatct
     6061 gtgtaatctt ttttacctgt gaggtctcgg aatccgggcc gagaacttcg cagcgagctc
     6121 attgtaccgc gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6241 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     6301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6361 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6421 tggaggccta gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt
     6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7261 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8581 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8641 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     8701 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     8761 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     8821 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg ccaagtacgc
     8881 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     8941 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     9001 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     9061 gtctccacc
//
