LOCUS       sequence-393636-        6793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid contains the TG1 int/attP locus and the 
            apramycin-resistance gene (aac(3)IV) for selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  AUTHORS   Gao H, Murugesan B, Hossbach J, Evans SK, Stark WM, Smith MCM
  TITLE     Integrating vectors for genetic studies in the rare Actinomycete 
            Amycolatopsis marina.
  JOURNAL   BMC Biotechnol. 2019 Jun 4;19(1):32. doi: 10.1186/s12896-019-0521-y.
  PUBMED    31164159
REFERENCE   2  (bases 1 to 6793)
  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: "BMC 
            Biotechnol."; date: "2019-06-4"; volume: "19(1):32. doi"; pages: " 
            10.1186/s12896-019-0521-y"
COMMENT     Sequence Label: pHG4
FEATURES             Location/Qualifiers
     source          1..6793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..60)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(44..60)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(57..79)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    68..84
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(92..98,99..116,117..122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    137..158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(275..292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(446..1034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(526..545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1205..1996,1997..2065))
                     /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     1828..1847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2066..2170)
                     /gene="bla"
                     /label=AmpR promoter
     oriT            3427..3536
                     /note="incP origin of transfer"
     CDS             3852..4652
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     protein_bind    4662..4708
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(5664..5675)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 gccgtgacgc ttactctaga gtcgaaagct tggcgtaatc atggtcatag ctgtttcctg
       61 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      121 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      181 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      241 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      301 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      361 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      421 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      481 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      541 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      601 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      661 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      721 ccgaccgctg cgccttatcc cgtaactatc gtcttgagtc caacccggta agacacgact
      781 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      841 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      901 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      961 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1021 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1081 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1141 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1201 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1261 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1321 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1381 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1441 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1501 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1561 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1621 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1681 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1741 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1801 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1861 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1921 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1981 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2041 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2101 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2161 gggttccgcg cacatttccc cgaaaagtgc cacctgacta gtccactgcc tctcggtaaa
     2221 atccagcaaa aattaatcag tgcagctcgc tgcactgatt aatttttgat caataggaga
     2281 tcgcttgtga cggcaagcac attgaaatct gttgagtagg cctgttattg tcgcccccag
     2341 gagacggaga atctcgacgg gggcgcagat gagattcaac ttattgggac gtgtccatgt
     2401 aatcaccgat gcgggatgtg taattccgct taaatcctcg aaggcgaccc agctcctggt
     2461 gctgctgctc ctcaggcggc acgaggtggt gggatcgggg gtgctcatcg aggagttgtg
     2521 ggcggaccac ccgccccgca gcgccatgac gacgctgcag acgtacgtgt accacacccg
     2581 ccggctgctg ggggagcacc gggtgacgag cgacgaccgg gaattggtcc tgacccagcc
     2641 gcccggctac ttcgccctga tcgacgagga cgaactcgac gtcgcggtcg ccgagcgtct
     2701 gatccgcacc ggcggccggc tgctcgagga gaaccggctc gaggaggcgc tcgcctcgtt
     2761 ggacgcggga ctggatctct ggcgaggccc ggcgctgtcc accgtaccgt gcggccgggt
     2821 gctcgaaagc aatatcgcgc acctggaaga gctgcggctt tttggaatgc agctccgtat
     2881 cgacgcgaat tggcggctgg gcagaatagg gccgatgatt ccggaactcc ggtccctggt
     2941 aatttcgcat ccgctgaacg agaccctgca cgccaaactg atgggcgcgc tctgtcagat
     3001 gggcaggcgc gccgaggcgc tggaatcgta tcggaatctc cggcggatac tgtccgacga
     3061 actgggggtg gatccgacgc cggaaatcca gcgtatgcac atggaaattc tcaacggtga
     3121 gaaggtgctc gtgtagcacc ggtccgtgaa cgcggtggag cccctcttaa gtgttcccct
     3181 ccctgcctcg tggtccctca cgcgctcagc tttgggcgcc cggctcgagc ggcggtcgaa
     3241 gggagatggg gtgccgctgg acgcggcgcc ggtggatccg gcatcgaggg gtcccgtatc
     3301 ggccttcgag cctccttcga gccacggggc cgacgatgac gacgaccacc ggacgaacgc
     3361 atcgattaat taagcggccg ccatatggaa ttcggtaccg catgcagatc taggaacttc
     3421 gaagttcccg ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag
     3481 ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct
     3541 gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc
     3601 gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg
     3661 cagcggaaaa tgcagctcac ggtaactgat gccgtatttg cagtaccagc gtacggccca
     3721 cagaatgatg tcacgctgaa aatgccggcc tttgaatggg ttcatgtgca gctccataag
     3781 caaaagggga tgataagttt atcaccaccg actatttgca acagtgccgt tgatcgtgct
     3841 atgatcgact gatgtcatca gcggtggagt gcaatgtcgt gcaatacgaa tggcgaaaag
     3901 ccgagctcat cggtcagctt ctcaaccttg gggttacccc cggcggtgtg ctgctggtcc
     3961 acagctcctt ccgtagcgtc cggcccctcg aagatgggcc acttggactg atcgaggccc
     4021 tgcgtgctgc gctgggtccg ggagggacgc tcgtcatgcc ctcgtggtca ggtctggacg
     4081 acgagccgtt cgatcctgcc acgtcgcccg ttacaccgga ccttggagtt gtctctgaca
     4141 cattctggcg cctgccaaat gtaaagcgca gcgcccatcc atttgccttt gcggcagcgg
     4201 ggccacaggc agagcagatc atctctgatc cattgcccct gccacctcac tcgcctgcaa
     4261 gcccggtcgc ccgtgtccat gaactcgatg ggcaggtact tctcctcggc gtgggacacg
     4321 atgccaacac gacgctgcat cttgccgagt tgatggcaaa ggttccctat ggggtgccga
     4381 gacactgcac cattcttcag gatggcaagt tggtacgcgt cgattatctc gagaatgacc
     4441 actgctgtga gcgctttgcc ttggcggaca ggtggctcaa ggagaagagc cttcagaagg
     4501 aaggtccagt cggtcatgcc tttgctcggt tgatccgctc ccgcgacatt gtggcgacag
     4561 ccctgggtca actgggccga gatccgttga tcttcctgca tccgccagag gcgggatgcg
     4621 aagaatgcga tgccgctcgc cagtcgattg gctgagctca taagttccta ttccgaagtt
     4681 cctattctct agaaagtata ggaacttcga agcagctcca gcctacacgc taggaacagt
     4741 tgcttagggc aacgaaccga cgccgggcac gagaaaaggg gccgaagccc cttcccgttg
     4801 cgctggtgcc gccgtcacgc cgccgctgtg aacccgttca gcgtctccgg ctccgtctct
     4861 tcctcgacct tcggcttcgc ccagtggata cgcacacggt cggcgatcgg gggaacccgt
     4921 ccgggccgct gaacgccctt cggaagcttg atcacctcga tccggtccac gaagagccgg
     4981 acgaacgccc gacggtcttc aaggggcgct agcgcccacc acgaacccgg tcccgtcggg
     5041 tcgctgcccc ggtcgcccag ccattcgccg atcggcagca cgggggagtc ggcggcgtcg
     5101 agctgacgaa gccgttcttc ggccccttcc atgcggagcg tcagcgcggc ttcctcttcg
     5161 agaaacgcgc gccgtcccat ggcggaccgg tagccgccgt tccgcttgtc ttcgtaaagc
     5221 tcttcgagcg ccttcacggc gtccgcgcgc tgtgccatca gttcggagcg ctgacccttc
     5281 aactccgggc gttcgtgcgt cgctccccag cgccgtgcgg cttcgtacat gagcgctgcc
     5341 gtgtcgcctt cgaggtcgtc agggtcggca gggtcgaagg ccgtgatgcg cgcgaagatc
     5401 gcgccgacga tgtacgggtc aaggttgtgc atcgtgatcg agcatgacga cccgtcatga
     5461 acccggcggg ggcacttgca cgcgtacgac gacttgtgag cttgatcacc ttcgcggacg
     5521 ttgcccgcca tggtcgaccc gttgctgtaa cccgtctccg ggtcgagctg gccggagccg
     5581 tagcagtaaa ggacgtccat tgccgacagg agcgattgcc cccggtactg acccttcccg
     5641 cgtcctcgac cctgaagcca ctcctgaagt tcccaccatt ccgccggggg aatgtacggc
     5701 tcgaagccgg gcagggtcag cggctccatg gtgaccgggt cgcgcctgat cttgtaatgg
     5761 ctgaagccgc cccgcgaacc gtcggcgcgc accttgtatg cgatgtcagc ttgatacccg
     5821 gcaatgcgcg ggtcgctgag tacgcgcttc aaaacgccgg ggtcccaatc ggaaccggcg
     5881 cgcttcttac cgaccagcgt gccgcgcgta ggcaccttgt cgcggtacag ccgctcacaa
     5941 agcccgttca gcgacccagg gtgaaacgac ccggctccgc cacccttgaa gggcgtatcg
     6001 cggtgcgtct tgatctcctg ccacgcccag cggattaccg ccccttcgct gccgtgtgcg
     6061 ccttcccagg tgtgcgcgct gggcaccagt cggcgaatgg caaccagctt tccgccgtct
     6121 tccgggttcg gaaccatttc ctcgaccgtg tcgaatccgt acggcgtcga ccccgtgtgt
     6181 ccgcccagcc gcttcgccaa ttccttagcg ttcgacacgg cgacgctctt gttcttcgac
     6241 tcatcatgcg aagcctgaag gcgcatgatc aggtgaataa ggtccatcat ttcgccgggg
     6301 cggaatgtgc cttcgttcac gctgacaatg gtcacgccca gccggagcaa ttccgtgacg
     6361 accggaataa tgtccagcgg ctcttcgcgg ctgaggcgcg aaatgtaatg aacaatgatc
     6421 atgttcattt cccggttccg gcacatgtcc aaaatccggt tgaactccgg ccggtcgacg
     6481 cccgtgaatg ccgacgtgcc gggcgcttcg ctgaagtgac ccagccactt cacctcgacg
     6541 ccgtcgcgcg cgtactcctt cgccagcgcc tcagccttcc cccggttcgc ggcgcgctgg
     6601 gtggccggtg aagcggtcga actgttctcc cgttccgccg actgtcggtc gtagccgcct
     6661 gccagaatga ccatgccttc aaccttcacg ctcatgccgg gcaggctatc ccgcccaact
     6721 gggtaagagc aaagactaac actgttgggc tggaacgggg ttgacctgcg gaaacgtctc
     6781 tcagtcggcg cgt
//
