LOCUS       sequence-215838-        2927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  efyp in pStA0.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2927)
  AUTHORS   Taylor GM, Mordaka PM, Heap JT
  TITLE     Start-Stop Assembly: a functionally scarless DNA assembly system 
            optimized for metabolic engineering.
  JOURNAL   Nucleic Acids Res. 2018 Nov 20. pii: 5193345. doi: 
            10.1093/nar/gky1182.
  PUBMED    30462270
REFERENCE   2  (bases 1 to 2927)
  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: "Nucleic 
            Acids Res. 2018 Nov 20. pii: 5193345. doi: 10.1093/nar/gky1182."
COMMENT     Sequence Label: pStA0::eyfp
FEATURES             Location/Qualifiers
     source          1..2927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..95
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             join(112..114,115..117,118..831)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(157..178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(418..437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     765..786
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      complement(861..947)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(960..978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1046..1150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1151..1219,1220..2011)
                     /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     complement(1369..1388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2182..2770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2671..2690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
       61 tcggttgccg ccgggcgttt tttattggtg agaatccagt gtggctcttc gatggtgagc
      121 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      181 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      241 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      301 accttcggct acggcctgca atgcttcgcc cgctaccccg accacatgaa gctgcacgac
      361 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      421 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      481 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      541 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
      601 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
      661 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
      721 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
      781 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta atgaagagcg
      841 taagacctct agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      901 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cccgaaaggg
      961 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1021 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1081 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1141 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1201 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1261 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1321 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1381 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1441 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1501 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1561 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1621 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1681 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1741 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1801 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1861 gcgtgggtcc cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1921 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1981 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2041 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2101 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2161 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2221 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2281 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2341 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2401 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2461 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2521 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2581 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2641 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2701 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2761 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2821 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2881 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgccttg
//
