LOCUS       sequence-233112-        6003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible EYFP on pAL5000/pMB1 backbone with kanamycin 
            resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6003)
  AUTHORS   DeLorenzo DM, Rottinghaus AG, Henson WR, Moon TS
  TITLE     Molecular Toolkit for Gene Expression Control and Genome 
            Modification in Rhodococcus opacus PD630.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):727-738. doi: 
            10.1021/acssynbio.7b00416. Epub 2018 Feb 5.
  PUBMED    29366319
REFERENCE   2  (bases 1 to 6003)
  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: 
            "10.1021/acssynbio.7b00416"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "727-738"
COMMENT     Sequence Label: pDD56
FEATURES             Location/Qualifiers
     source          1..6003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(106..694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(186..205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(978..1772)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1089..1108)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1699..1718
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3672..4550)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4577..4861
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4784..4803
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(4898..4900,4901..4903,4904..5614)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4943..4964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5204..5223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5551..5572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5649..5720
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5737..5808
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    5809..5829
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5830..5887
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5911..5928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
       61 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      181 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      301 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      541 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      661 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      721 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      781 aaggatcttc acctagatcc ttttggggtg ggcgaagaac tccagcatga gatccccgcg
      841 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
      901 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
      961 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     1021 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     1081 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     1141 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     1201 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tccgcgcctt gagcctggcg
     1261 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     1321 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     1381 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     1441 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     1501 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     1561 gccagccacg atagccgcgc tgcctcgtct tggagttcat tcagggcacc ggacaggtcg
     1621 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     1681 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     1741 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     1801 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     1861 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct
     1921 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
     1981 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt
     2041 cagcaccgtt tctgcggact ggctttctac gtgttccgct tcctttagca gcccttgcgc
     2101 cctgagtgct tgcggcagcg tgaagctagc gtctaccagg acttttacct gtccgacccg
     2161 ttgcaacgga accccccacg gaacccccgc gacacccgct ccccaattgc gttagaacag
     2221 cggtggattg tcggcttcgt tgtgggcctt ttgagccgct tcctgttctg ccgcacgctc
     2281 tttcctcgcc cgatagccga gtcgcttaac ggtgtccaga tgcagcccga aatgtttggc
     2341 cgtttgcggc caagagtggc cctcgtcgtc gtgataggcg cggatgcgtt cgcggcgtgc
     2401 agcctgctcg gcgagccact cgctgcgttc ctgcgccacg agccggacga cgtggcgttc
     2461 ggatagtccg gtgattcgag cgccttcggc ggcggtcacg cgccgctttt tgcggacagt
     2521 cggctgccgg ttgtagccgt cgctgtagcc gtcgctcata gcaatgcctc catggctgac
     2581 gcggactttg cgcgccgcgc aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc
     2641 gagatggccg actggcgcgc actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa
     2701 atgcgcgact tggttgtgat ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg
     2761 tccggtagcg gtccgggaca cacgtcgttg cacgggaact cggcgtttcg cgcgtggcac
     2821 tcggcataga tcgcgcggcc gagtccgtcc acgttccggg tcggcaggta gatccgcatg
     2881 agggcgggac gataggccca caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc
     2941 ggcgtcggag ccgctttgta cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg
     3001 ccgagctcgg cctcgatgtg gctgagtgtg tagagatctg agtggagcca ttccgtttcc
     3061 caggcgatgt ggccggggtt tttggtcatg aggcctgagt aactgcggtc gccgtccacg
     3121 gcgcgccgaa ggccttcggc gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat
     3181 tcggtgcgtg gaacaggggc gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga
     3241 ttgcccacga tcgcgttggg cagcggatgg gacccccggg cgctgagcgc tcggagcgct
     3301 gcgtctggat ggtctacgtc cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg
     3361 atgtaccggc ggcctagggc cgacgcgcgg ctttggcggt agatcccctc gagcagatcg
     3421 tcgcttgcca gcggccagta cggcagccag agctgctcaa attcgtcggc gacgtggctc
     3481 acgcttggta gtagaccacg attaatcacc ggtgtatggt ccgacacgag ctccaagtca
     3541 gatatttcgc tgaggggcca ccccacaact gcacactccc ccgctctccc gtcgagccct
     3601 ggtggtggaa caccagcgac agccgagcac ccccaaccac ctgtaccaac cactagttga
     3661 ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
     3721 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga
     3781 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
     3841 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
     3901 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
     3961 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
     4021 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
     4081 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
     4141 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
     4201 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
     4261 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     4321 cctggcggga acagtaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     4381 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     4441 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     4501 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     4561 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
     4621 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
     4681 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
     4741 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
     4801 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
     4861 tacagcggat aaagtagcaa agagaaggag gttaggaatg gtgagcaagg gcgaggagct
     4921 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4981 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5041 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     5101 cctgcaatgc ttcgcccgct accccgacca catgaagctg cacgacttct tcaagtccgc
     5161 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     5221 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     5281 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     5341 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5401 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     5461 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     5521 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     5581 cgggatcact ctcggcatgg acgagctgta caaggctgcg aatcaaaaat aatactagag
     5641 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     5701 gtttgtcggt gaacgctctc tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg
     5761 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagtagcgg
     5821 ccgctgcagt ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt ctttaaaacc
     5881 gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt
     5941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6001 agg
//
