LOCUS       sequence-397088-        5319 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 5319)
  AUTHORS   McLaughlin RW, De Stigter JK, Sikkink LA, Baden EM, Ramirez-Alvarado
            M
  TITLE     The effects of sodium sulfate, glycosaminoglycans, and Congo red on 
            the structure, stability, and amyloid formation of an immunoglobulin
            light-chain protein.
  JOURNAL   Protein Sci. 2006 Jul;15(7):1710-22. Epub 2006 Jun 2.
  PUBMED    16751605
REFERENCE   2  (bases 1 to 5319)
  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: "Protein 
            Sci."; date: "2006-07"; volume: "15(7)"; pages: "1710-22. Epub 2006 
            Jun 2"
COMMENT     Sequence Label: pET12a-AL-09 FL
FEATURES             Location/Qualifiers
     source          1..5319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..99
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        80..98
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(924..942)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      938..985
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1351..1356,1357..1373,1374..1379))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1406..1424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1492..1596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1597..1665,1666..2457)
                     /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(1815..1834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2628..3216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3117..3136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3370..3387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3402..3541
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3467..3486)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3627..3649
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3643..3834)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5296..5315
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
       61 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      121 agaaataatt ttgtttaact ttaagaagga gatatacata tgcgggcgaa actcctagga
      181 atagtcctga caacccctat cgcgatcagc tcttttgcgt cgacggacat ccagatgacc
      241 cagtctccat cctccctgtc tgcatctgta ggcgacagag tcaccatcac ttgccaggcg
      301 agtcaggaca ttaacaacta tttaatttgg tatcagcaga agccagggca agcccctaag
      361 ctcctgatct acgatgcatc cactttggaa acaggggtcc catcaaggtt cagtggaagt
      421 ggatctggga cagaattcac tttcaccatc agcagcctgc agcctgagga tcttgcaaca
      481 tatcactgtc aacagtatga taatctcccg tacacttttg gccaggggac caagctggag
      541 atcaaacgaa ctgtggctgc accatctgtc ttcatcttcc cgccatctga tgagcagttg
      601 aaatctggaa ctgcctctgt tgtgtgcctg ctgaataact tctatcccag agaggccaaa
      661 gtacagtgga aggtggataa cgccctccaa tcgggtaact cccaggagag tgtcacagag
      721 caggacagca aggacagcac ctacagcctc agcagcaccc tgacgctgag caaagcagac
      781 tacgagaaac acaaagtcta cgcctgcgaa gtcacccatc agggcctgag ctcgcccgtc
      841 acaaagagct tcaacagggg agagtgttag taaggatccg gctgctaaca aagcccgaaa
      901 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      961 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atccacagga
     1021 cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga
     1081 ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca
     1141 tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga
     1201 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
     1261 tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc
     1321 gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa
     1381 catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     1441 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1501 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1561 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1621 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1681 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1741 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1801 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     1861 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1921 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1981 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2041 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2101 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     2161 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2221 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2281 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2341 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2401 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2461 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2761 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3241 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3301 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3361 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3421 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     3481 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     3541 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3601 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3661 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3721 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3781 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     3841 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     3901 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3961 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     4021 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     4081 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     4141 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     4201 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     4261 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     4321 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     4381 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     4441 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     4501 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     4561 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     4621 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     4681 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     4741 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     4801 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     4861 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     4921 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4981 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     5041 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgatcg
     5101 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     5161 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     5221 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5281 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcc
//
