LOCUS       sequence-95474-e        4605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses AmiE with proK14 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4605)
  AUTHORS   Bienick MS, Young KW, Klesmith JR, Detwiler EE, Tomek KJ, Whitehead 
            TA
  TITLE     The Interrelationship between Promoter Strength, Gene Expression, 
            and Growth Rate.
  JOURNAL   PLoS One. 2014 Oct 6;9(10):e109105. doi: 
            10.1371/journal.pone.0109105. eCollection 2014.
  PUBMED    25286161
REFERENCE   2  (bases 1 to 4605)
  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: "2014-10-6"; volume: "9(10):e109105. doi"; pages: " 
            10.1371/journal.pone.0109105. eCollection 2014"
COMMENT     Sequence Label: pJK_proK14_amiE
FEATURES             Location/Qualifiers
     source          1..4605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(560..1351,1352..1420))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1183..1202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1542..2130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2031..2050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2284..2301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2316..2458
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2384..2403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2544..2566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2560..2751)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     RBS             3390..3412
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4449..4466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4519..4537)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4533..4580
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct taccaatgct taatcagtga ggcacctatc tcagcgatct
      601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
      781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
      841 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
      901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat aacacccctt gtattactgt
     1441 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     1501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1681 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     2341 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     2401 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     2461 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2521 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2581 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     2641 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     2701 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     2761 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     2821 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2881 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     2941 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     3001 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     3061 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3121 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3181 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggccacagc
     3241 taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg gataactgta
     3301 cgggcatgca taaggctcgt aatatatatt cagggagacc acaacggttt ccctctacaa
     3361 ataattttgt ttaactttct agaaataatt ttgtttaact ttaagaagga gatatacata
     3421 tgagacatgg cgatattagc tcgtcaaatg ataccgtagg cgtagccgtg gtgaattaca
     3481 agatgccgcg tttacatact gctgctgaag tcctggataa tgcccgcaaa attgcggaaa
     3541 tgatcgttgg tatgaagcaa ggtctgccgg gcatggatct ggttgtgttt cctgaatatt
     3601 ctttacaggg tattatgtac gaccctgctg aaatgatgga aacagccgtg gcgattccag
     3661 gcgaagaaac ggaaatcttt agccgtgctt gtagaaaagc aaatgtttgg ggtgtgttct
     3721 ccctgaccgg cgaacgtcat gaagaacacc ctagaaaggc accatacaac actctggtct
     3781 tgatcgataa caacggtgaa atcgtacaaa agtacagaaa gatcatccca tggtgtccga
     3841 ttgaaggctg gtatccaggt ggccagacat acgtctctga aggtccgaaa ggcatgaaga
     3901 tctcattaat tatctgcgat gacggtaatt atccggaaat ttggagagat tgtgccatga
     3961 agggtgcgga attgatcgtt cgctgccaag gctatatgta ccctgctaaa gaccaacaag
     4021 ttatgatggc taaggcaatg gcctgggcga ataactgtta tgtcgctgta gcaaacgctg
     4081 caggttttga tggcgtttat agctacttcg gtcatagtgc cattatcggt tttgacggcc
     4141 gtactctggg tgaatgcggc gaagaagaaa tgggcattca atacgcgcag ttgtctctgt
     4201 cacaaatccg cgatgcccgt gcgaatgacc aaagtcagaa ccatttgttt aaaatcttgc
     4261 acagaggtta ctccggtttg caggcttcgg gcgatggcga ccgtggtctg gcagaatgcc
     4321 catttgaatt ctaccgtacc tgggttactg atgctgaaaa ggcaagagaa aacgtggaac
     4381 gcctgactcg ctccacaaca ggtgtcgccc aatgcccagt aggtcgtctg ccgtatgaag
     4441 gcctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     4501 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     4561 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggat
//
