LOCUS       sequence-41829-e        5128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5128)
  TITLE     pHyVec Cloning Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5128)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pHyVec4
FEATURES             Location/Qualifiers
     source          1..5128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1714..2430
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1744..1772)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2367..2387
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2940..2960)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2940..2958)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2979..2995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2979..2995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2992..3014)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3003..3019
                     /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(3027..3033,3034..3051,3052..3057))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3072..3093
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3210..3227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3381..3969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3461..3480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4140..4931,4932..5000))
                     /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     4763..4782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5001..5105)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggttttaag
      661 ccgagatgca ggattctgaa tgaaaaagaa aaaaagaagt ctcggtagag taaaagtgat
      721 cggtttgcaa ctgtaaaatt tattgaagta ccaataattt tatttaaaat aaaactgaaa
      781 tataaagtta aagttgctgt tctataagtt taccgaattt taaaaccatt gtaacgctag
      841 agtaatattt gagtctacta agttagtccc cgcacttttt aatcaagcaa taaataccca
      901 aactttgctt attcaaatca ataaaccaat atatctctta aaataaagta aaaacttctg
      961 aaattctata aaaaaaaatt taatttcgaa atatcaaatg taacttcaac accgcactat
     1021 tttcttttaa acaactgata tagtaattac ttctcaaaaa cgttatctca aggtttgtga
     1081 tgtacttaaa accactccta ttttgttacg cgtttaaaaa agcaaacata agttggtttc
     1141 tattgatgaa tgagaacata tttcatttaa agttaaaatc ctaccagtgg tttcactgta
     1201 cgtaaacacc gtcaaaaaaa caggaacgtt tttaaagatt aataattgaa gtaaaaaaaa
     1261 tttaataccg ggggttaaaa aaatctttta aaataattat aaatatatat attaaaattt
     1321 ataaattttt aaacacattt aaaatatata ttaagtataa taaaagtaat attataaaaa
     1381 aaaatttaat tttataatta tttttattaa atttataaat aataggtaaa acttacatat
     1441 ccgttttatt ttttcttaat aaaataacgc gtgcaaattt ttgtccatat aaagaccttt
     1501 tcgaacaata acttttttgc ttagccgttt tttttcttat atggtcaaaa aagcgctcaa
     1561 gcgattcacc ataaaaagcg caattagttc agcgttcgtt attcagaagc ttcagctttg
     1621 cttgatactc agctcttctc tttttaaaca aaacactaat caagctagcc gatgatgaag
     1681 ttgcccgccc tcgctgcagc acccggggaa aaaatgagta aaggagaaga acttttcact
     1741 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     1801 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
     1861 actggaaaac tacctgttcc atggccaaca cttgtcacta ctttctgtta tggtgttcaa
     1921 tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc
     1981 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
     2041 gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat
     2101 tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat
     2161 gtatacatca tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac
     2221 aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc
     2281 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa
     2341 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt
     2401 acacatggca tggatgaact atacaaatag cattcgtaga attcacaatt cgattatatt
     2461 tatactggac tatttttaca tctgttcggt tattttcaca tttatttttc tatatatatc
     2521 ttataaacgt tttaaaaccc atgtaatttt tgttaagctg taatataaaa gacgtcctaa
     2581 caaacttctt ttattactga atttccttta attataataa ataacaagtt ttaaaataaa
     2641 ttcaggcaat taaggcgctc ctgaggtact aaaattaatg taaacattta aaattaactt
     2701 ggatggtctt aagtactgta ctcgtgattt tgttatactt tattattaga aaagtcgtct
     2761 attaactttt tgttccttaa tttacttgat taaattgtcg cttaatttat caaatcaggt
     2821 tttgcgcgtt attttagaga aaaacttatt agaaaaatga ataagcaaag tttaggctaa
     2881 catgtttttt tattatttta aatagttcaa gtcaatgacg tataaaccag cttttgttcc
     2941 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     3001 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3061 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3121 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3181 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3781 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5101 ccgcgcacat ttccccgaaa agtgccac
//
