LOCUS       sequence-93646-e        3748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSALECT with Efnb2 insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3748)
  AUTHORS   Hsiau TH, Sukovich D, Elms P, Prince RN, Strittmatter T, Ruan P, 
            Curry B, Anderson P, Sampson J, Anderson JC
  TITLE     A method for multiplex gene synthesis employing error correction 
            based on expression.
  JOURNAL   PLoS One. 2015 Mar 19;10(3):e0119927. doi: 
            10.1371/journal.pone.0119927. eCollection 2015.
  PUBMED    25790188
REFERENCE   2  (bases 1 to 3748)
  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: "PLoS One.";
            date: "2015-03-19"; volume: "10(3):e0119927. doi"; pages: " 
            10.1371/journal.pone.0119927. eCollection 2015"
COMMENT     Sequence Label: pSALECT-Efnb2
FEATURES             Location/Qualifiers
     source          1..3748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             698..1486
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(844..863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(1790..2449)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2364..2383
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2450..2552)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2772..3360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3261..3280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3514..3531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3648..3669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3684..3689,3690..3707,3708..3714)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ggatctcgac atgaacaata acgatctctt tcaggcatca cgtcggcgtt ttctggcaca
       61 actcggcggc ttaaccgtcg ccgggatgct ggggccgtca ttgctaacgc cgcgacgtgc
      121 gactgcgcat atggaattca tgcggtccat cgttttggag cccatctact ggaacagcag
      181 taattctaag tttttaccgg gccagggcct tgtgctctac ccgcagatag gggataaact
      241 agacataatt tgcccgaagg tcgattcgaa aacagtcggg caatatgaat actacaaagt
      301 atacatggta gataaagatc aagcagatcg ttgtactatt aagaaggaga acacaccgct
      361 gctcaactgc gcccgaccag atcaagacgt gaaattcacc atcaaattcc aagagttctc
      421 tcccaattta tggggattgg agtttcaaaa gaacaaagat tattatatca tttcgaccag
      481 caatggcagc ttagaagggc tagacaacca ggagggtggt gtttgccaga cgcgtgcaat
      541 gaagattctc atgaaagtgg gccaggatgc cagctctgct gggtctgctc gtaatcacgg
      601 tccgacacgc agaccggaac ttgaggccgg gacgaacggc cgttcctcca ctacaagccc
      661 tttcgtgaaa cctaatcctg gtagctccgg atcccaccca gaaacgctgg tgaaagtaaa
      721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
      841 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
      901 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
      961 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1021 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1081 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1141 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     1201 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1261 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1321 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1381 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1441 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaagatt tttccaatga
     1501 tgagcacttt ttgatcagaa aaaaaggatc atatcgtcaa ttattacctc cacggggaga
     1561 gcctgagcaa actggcctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     1621 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     1681 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     1741 aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc
     1801 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     1861 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     1921 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     1981 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     2041 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     2101 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     2161 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     2221 ttgccatacg taattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     2281 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     2341 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     2401 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     2461 tagctcctga aaatctcgac aactcaaaaa atacgcccgg tagtgatctt atttcattat
     2521 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc
     2581 agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg
     2641 tcacaggtat ttattcggtc gaaaaggatc taggtgaaga tcctttttga taatctcatg
     2701 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2761 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2821 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2881 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2941 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3001 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3061 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3121 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3181 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3241 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3301 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3361 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3421 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3481 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3541 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3601 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     3661 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     3721 attgtgagcg ggataacaat ttcacaca
//
